And welcome back to a special edition of the Falcons audible presented by AT&T. I'm Derek Rackley and I'm back with my fellas DJ Shockley and Dave Archer. And yes, it is a special edition because the top two spots at least as far as football is concerned have been filled by the Atlanta Falcons. Arthur Smith has come over as the offensive coordinator previously with the Tennessee Titans and he now is the 18th head coach in Falcons history for the Atlanta Falcons. And then the same day it was finalized that Terry Fontenot comes over from the New Orleans Saints, a longtime executive in the Saints organization. He will take over as the general manager role and those two will work side by side to build this organization back up to what the owner Arthur Blank wants and feels like the fans of Atlanta deserve. So we will talk a lot about what's going on with those two guys, the whirlwind that they're going through in their new positions. But real quick before we get to the guys, I want to make sure I tell you what we've got in store. Again, reactions to the top two guys for the Atlanta Falcons. We'll talk about how is Arthur Smith going to duplicate what he did in Tennessee without some guy named Derek Henry because that was a pretty integral piece to the puzzle that he had up in Nashville. And we'll talk about the general manager Terry Fontenot as he comes over and he starts to analyze this roster. What are some of his first moves as a member of the Atlanta Falcons organization? We'll talk a little bit about free agency in the draft and how those two guys end up working together to build this roster up to where they feel comfortable to where week one, they're ready to be competitive and go out and win football games. And then of course we'll just kind of look into our crystal ball a little bit, look out in the future and see where we think things are going to go for the Atlanta Falcons. So with that being said, let me get to the fellas here because DJ and Dave, they want to talk, they like to talk and they've got good things to say when they do talk. Fellas, it's been a little while, we've had some playoff action. So DJ, let me start with you. Let me get your first impressions when you heard of Arthur Smith being hired and Terry Fontenot coming over as the head coach now and the general manager, those two positions. Well, when you first hear about it, obviously we, we talked about it some weeks back about the candidates that can possibly roll into this spot and during that time, that's when I started to familiarize myself with a couple of these guys and wondering, okay, if this guy comes to Atlanta, what does it look like? If this guy comes, you know, where's this guy been? What has he done? And we finally settled on Arthur Smith and Terry Fontenot and we got a chance to hear their press conferences. And one thing I took away immediately was these guys both have an understanding of who they are first off and what they want to do going forward. I was impressed by both guys. I thought both guys handled the Atlanta media for the first time really well. I thought they were asked a couple of hard questions that were questioning. I'm sure a lot of fans wanted to know, but I thought both guys were really sounding what they wanted to do. And a couple of things that stood out to me when Arthur Smith was talking and a lot of it was about the identity and how he came to this spot and his dad, all that kind of stuff. But the one thing he talked about was being great up front. And I think that's exactly where it starts in the National Football League. I think it starts here in college. I think it starts here in high school. And that's the number one thing that he talked about is he's a guy who's played it. He's played offensive line. He's talked offensive line. And the fact that he's been there and done that and has a where for how to teach these guys, I think was big. And then the one thing I took away from Terry, which was really, really good was him talking about his willingness to listen, learn and adapt. And I think as a GM, that speaks volumes because you come in here and you have everything at the same time. You can do this, you can do that. There's so many different ways you can go. But the fact he says, Hey, I got to have people around me who are smart to me. I'm not going to have people around me who just going to be yes people, but their willingness to listen, learn and his ability to adapt, I think was huge. And some of the things he talked about. So. The other thing that really struck me was he talked about sustained success. And I know that's probably one thing that a lot of Falcons fans are going on right now. We're going out what's going on for the future. And I think sustained success means right now as well as the future. And I love to hear that part of it. So I was truly impressed by both guys, really bright guys. I can see why all the money rich McKay were really excited to have these guys a part of it. And Hey, Terry was the first one they interviewed. And Terry fun to talk about, Hey, I went out and I interviewed these guys myself before even Arthur them said anything to me about it. That tells me he has the initiative to go out and see what he likes about these coaches. So I thought that was impressive as well. Yeah. DJ, a bunch of great points that you make there as we get the dogs in the background that want to chime into there'll be your time as well. Okay. There will be your time to get the dogs into the show. You'll Dave, I think the interesting point here though is that the Falcons have two guys coming into this position that have not been in that position before they have not been head coaches. They have not been a general manager before, which that's not always the case in the NFL.  So what were your first impressions after seeing Arthur Smith and Terry font know in their press conferences today? Well, the first thing that struck me is these are two guys that are very humble and DJ talked about listening. I think that's why they've risen in the ranks of where they are is because they're very humble. They're very thankful for their opportunity. I think that's something that Terry font knows said in his comments. He says I was my goal was not to be a GM in the National Football League. My goal was to do the best possible job with the opportunity I was given. He was also asked about how did you pitch yourself to the team? So I didn't pitch myself to the team. I told him who I was. These are my values. This is what my philosophy is. And I was very transparent of who I am. And if that fits with the Atlanta Falcons, which it did, then so be it. I think that's what I'm talking about. I think that's what I'm talking about. I think that's what I'm talking about in New Orleans. He's probably was the heir apparent to Mickey Luna. Loomis for the job in New Orleans. So he felt like the fit was perfect in Atlanta. He loved the ownership group and how do you not? I don't know. There's a better owner in the National Football League than Arthur blank. Another a better guy that's just under the owner than Rich McKay, a guy you can bounce stuff off of that's been around the league as long as.  They were in lockstep with a lot of these things. And you kept hearing about how they'd only known each other for a couple of days or talk to each other a few times. But the words that came out of their mouth were the same words. And so the vision came together very quickly as far as them being in lockstep as to how this thing is supposed to work. How you do anything is how you do everything. That stuck in my eyes. I said, wow, that, that's powerful. You know, that's not something that you hear all the time. And I know he has a, he has the ability to pull some quotes from some very, from very smart people, but that, that's a good one there. How you do everything, how you do everything. Or how you do anything is how you do everything. I thought that that was pretty impactful. You know, I've read some of the social media reaction on the news about Terry Fontenot and everybody was just putting their stamp of approval that he should be the guy for the job for the Atlanta Falcons. And we kind of all talked before we got on the podcast that we were all impressed with Terry Fontenot. It just seems like he's a very polished, knows what he wants. But to DJ's point also realizes that just because he's the boss, if you will, it's not that he has all the answers. He's going to lean on other people to give them insight, as he called it, layer of information, continue to deliver him layers of information on how he's going to end up help building this franchise and getting them competitive. I think it's interesting here, fellas, I wanted to point out, Terry Fontenot, when he talked about Mickey Loomis, as you mentioned, Dave, you got pretty choked up today. So you could see how important Mickey Loomis was in developing him, taking a chance on him as a marketing intern way back when, when he was at Tulane in college and growing him, up through the ranks teaching him basically everything that he knows. But Terry also said that we're probably not going to have a whole lot of conversations anymore as rivals in the same division, but he's not necessarily going to have that person to lean on. But Arthur Smith on the other hand, which is really intriguing. He will have somebody to lean on as his father is the founder of FedEx.  But how about picking up the phone and saying, Dad, I'm about to run this huge organization. Can you help me? You did a pretty good job at FedEx. You got a couple of tips for me. I mean, talk about that in psychosh. That would be phenomenal. So I want to, I want to kind of- And real quickly, Rack, let me add to what Frederick Smith brings to the table, the father of Arthur Smith. He's also a war hero. This is a guy that spent time in Vietnam, led men into battle. And so when you start talking, we always kind of compare sports to wrongly, by the way, but we compare sports to battle and or service in the armed forces. He actually was in Vietnam. Bronze star, silver star winner, led guys into battle, had to call in artillery strikes in a position where he was to help save lives, rescue a number of people. Yeah. You talk about leaning on a dude that built a multi-million dollar, billion dollar operation at FedEx. How about the fact that he's led guys into battle? That's a pretty good one to be able to lean on as well. Yeah. He's got some resources as far as making sure that he runs this thing from the top properly. And DJ, I wanted to pick your brain because let's talk a little bit about the action on the field and let's stay on Arthur Smith here because as soon as you heard about the news for Arthur Smith, you did a couple of breakdowns on what made him such an effective play caller for Tennessee. I think the biggest thing to note here, we talked a lot about it during this podcast throughout the course of the season was the lack of the running game, the presence in the backfield for the Atlanta Falcons. Now, Arthur Smith had it when he was in Tennessee. He had some guy that named Derek Henry that just rushes for over 2,000 yards and maybe one of the most feared running backs in the National Football League. So how does Arthur Smith do it in Atlanta without a guy in the backfield, at least as of right now, like Derek Henry? You know what? I had a chance to talk to Ben Jones, who is the starting center for the Tennessee Titans. And I kind of just picked his brain on what is Arthur Smith like, what is this offense like, and the things he does.  He talked to me about how the offensive line is looking at is it quarter, is it quarter quarter halves? If we got a hangover player, what they're doing here. And that is a direct correlation coming from the offensive coordinator and their coaches on how they prepare weekly. And yeah, Derek Henry is a star. Yes, we all know that he's a big physical downhill back. And yes, he's deserving of all the credit. But the more I went into the film, the more I dived in, there were times where Derek Henry wasn't getting touched until he was into the second or third level. And that's because of the way he created Run Game. And you're talking about formation. You're talking about motion. You're talking about using different shifts. You're talking about putting guys in position to get the right block. And we're talking about angles. We're talking about all these X's and O's kind of stuff. But it's simple stuff when you watch the film that allowed them to be one of the top rushing attacks in all of football. So when you talk about Arthur Smith, it's more to it than just Derek Henry. It's more to it than just Ryan Tannehill. The way he prepared his players week in and week out. And I went through five, six, seven different games. And it wasn't just one game they were doing. They were doing this every single week. So it shows you each week, they have the ability to adapt for one we talked about already, but to be able to change up what they do, but then find ways to create it for their players. And I think the one thing that he talked about is we're going to make sure we use our personnel to his best ability. And I think that's the number one thing is when some coaches come in, they say, all right, it's going to be my scheme. And this is what it's going to be about. The first thing he said is it's all about players, not plays. And I think that is the number one thing as a play caller. If you understand, I have a set amount of guys who can do this. I have a couple of guys who can do this. And then using those plays to fit those guys, I think it's tremendous. And Arthur Smith is a guy who uses a multitude of ways to create plays for his playmakers. And I saw it throughout the film. I saw it in the run game. I saw it in the pass game. I saw him change up personnel, change up formations. I mean, he does so many different things. And at the line of scrimmage, it's going to be fun to watch his team. I, I, I text Arch about it. And I say, you're going to be excited. I'm sure he's seen it through and through, but he's going to be excited to be able to call these games because of all the things that he does to give his guys a chance to win. And I just pre-snap, but post-snap. Yeah. And Arch, I think you can appreciate me here because, you know, you look at, uh, the Tennessee Titans offense. And I think Arthur Smith has been getting the credit for kind of reviving Ryan Tannehill's career by how he's been able to work with the personnel within that offense. Again, right now we're, the Falcons like they're not going to have a guy in the back field like Derek Henry, but maybe it's the creativity. And the other thing Arthur said in his press conference was adaptability, Dave. And, and speak to a little bit about how you feel like he needs to be more adaptable, maybe than the Falcons have been in the past to use some of the weapons that he has right now and some of the other ones that they're likely going to bring into this organization. Yeah. Let's look at what he did just from 2019 to 2020. That offensive line in Tennessee gave up 53 sacks in 2019. Okay. That was a shared quarterback duty between Marcus Marriota and with Ryan Tannehill. 53 sacks. This year they gave up 25 sacks. And that was with Taylor LaWan missing the better part of three quarters of the season with a blown ACL. And Tyson Brello. Yes. Tyson Brello had taken over at left tackle. He got hurt. So you had three different guys playing the left tackle position, their ability to adapt as you guys have talked about. And that's been an operative word. Is it an operative word that both these guys spoke about their ability to be adaptive to the personnel they had. To me, that speaks volumes, the ability to adjust that big group up front. He says we are standard along the offensive line is we're going to be great up front. And that's where you're, that's the brick and mortar guys of the house. If we could get after people physically, that helps the run game. That helps the passing game. Yeah. Nobody had, there's not a bunch of dairy kinders. In fact, there's one, but there's other people that run the football and there's other guys that could get that job done. If you're providing the opportunities like shock Ted said, the ability to prepare guys to know what they're looking at and make adjustments. Again, the word adapt to what you see based on strength of formation or personnel's outnumbering. Ryan being prepared to change the play to an, to an uncovered or outnumbered look to the other side to get the run game going. These are all things that, that are very exciting to me. And shock. And I both played the quarterback position to know that you're going to have options to be able to get to it, the line of scrimmage. So you're not running the football into a stacked front, or you can throw the football if they're trying to crowd you to line of scrimmage. Right. Right. Let me add one thing to what ours just mentioned, about the ability to do stuff at the line of scrimmage. I asked Ben Jones. I said, and this is two different things. He, he said, when he found out all this was going to be going, he immediately called him, text him or whatever it was and said, Hey, I know Alex Smith may be gone. I know, I mean Alex Mack, excuse me. He may have a chance to leave whatever it may be. Don't forget about me. I would love to come to play for you in Atlanta. But the second part of this was I asked him, who has the responsibility of the line of scrimmage? Is it you or is it a quarterback? How much of you guys work together? And this is his call. He said, we literally can change every single play at the line of scrimmage to whatever we want. If there is a situation where we're in a certain play and it doesn't fit or it doesn't look good, or it's going to be a negative play, the quarterback has an opportunity to change any play in the playbook at any time. And the freedom at the line of scrimmage for a quarterback is like gold. Arson's told you the ability to do that in line of scrimmage and not be caught in a play is tremendous. Well, and if, and if that ends up being the case and Matt Ryan is still the starter, you'd like to think Atlanta's going to be in pretty good shape because of all the experience and all the reps that he's had throughout his storied career would probably have a much higher up leg on a younger quarterback and being able to make all those adjustments to get the offense in the right play for them to have some success. Offensively. So we talked a lot about Arthur Smith.  And he's going to be kind of the orchestrator, if you will, as far as adding new pieces to the puzzle that are going to help Arthur Smith achieve what he wants to do offensively and for this entire team, defense and special teams, because they all talked about the three phases of the game and Arch, I want to talk with you about Terry Fontenau. He was very quiet today during his press conference on any personnel. He didn't feel like it was fair to be talking about personnel at this moment before he and whoever the rest of his staff ends up being have a chance to discuss it. So I can appreciate that, but let's put you and his shoes from a personnel standpoint. Give us one or two things. Give the fans one or two things that, that you as Terry Fontenau as the general manager would be taking a deep look at from this roster, from the personnel standpoint to help them improve in 2021. Well, there's a, I wrote down a couple of areas from a, from a personnel standpoint. Number one is probably the edge of your defensive front. What are you going to do there? What can you do? Obviously, there's some salary cap implications that might hamstring you a bit in free agency. There are a couple of players that potentially present themselves in the first couple of rounds of the draft that are intriguing. Another position is the safety position. You're going to have to make some adjustments there. We know Keanu Niels deals up. We know Ricardo Allen. You're going to bring Ricardo Allen back at safety. Demonte KZ is coming off of a blown knee. Are you looking to add a safety? I wrote down five safeties that are going to be available in free agency that might be affordable. How about the running back position? There's four or five running backs. Chris Carson, a local kid played high school football here. He's a guy in Seattle. Let's carry the mail up there. You've got a guy like that. That could potentially they're available. Then also we talked about potentially the last time we got together about maybe trading where you move back and garner some picks. There's some really good players in the draft. They're going to be available to address the running back issue. That's just a couple. The one question I have for you guys, and you got to help me out here. What are we doing at left guard? What's happening there? And is that a scenario where you see Jake Matthews slide inside and play left guard? Does he slide inside to play there? And now all of a sudden that number four pick is used on the big fellow from Oregon or, you know, just another thought as to what potentially number four could be. I know there's a lot of talk about quarterback in this. And Terry Fountain will talk about drafting and being able to stock up all strength. Hey, if there's a strength guy there, it's the best player available. Even though if we got a player there that may be playing for the next year or two, we can still draft the guy there. He's not going to shy away from that. But there's a guy sitting there that's supposed to be a generational left tackle that might be available. Could that help fix your offensive line? Yeah. And one of the things that he talked about is he kind of comes from that mindset of best player available and not necessarily reaching for a need, which we talked a couple of weeks ago about a guy named Greg Russo. I believe his name, the guy kid out of Miami. That's a pass rusher. But I think we all kind of agreed without saying it that at four, that's probably a little bit of a stretch, right? And so if, if now that Terry font knows the general manager, I don't think that that guy is in the mix as of right now. Granted, it's really early on. Maybe he goes and he dazzles during his pre draft prep, but he's not necessarily going to reach just because they need a pass rusher. But what happens if a Devonte Smith, if they feel like is the best player available at number four, they feel pretty good about wide receiver. Maybe he goes there just because his belief is best player available. So DJ, I'm going to ask you the same question. Personnel perspective, Terry font know now the general manager, what moves are you making to help rebuild this roster? I think arch made a bunch of great points and they are similar to some of the things I think about when I think about this roster, where do you head? Where do you go? Some key needs. One point that I think I would pose to our group and pose to fans as well is if you don't get that guy at four, you want or at four, you think you can move back. Maybe you get a couple of picks in that first round. Here's something to think about. Arthur Smith now is a guy who uses two tight ends a lot. There's a guy coming out from Florida named cow pits. We got Hayden Hertz. We also have an opportunity to get another tight end. He uses tight ends a lot. There's a lot of 12 personnel. There's a lot of 22 personnel. He uses tight ends a lot.  Well, here's another playmaker for you on the office side of the ball to go with these receivers as well. So you pose some different questions and like I said, they're just going to be options. Four is going to be a good spot.  But you're going to have an opportunity to move. You're going to have an opportunity to stay there, but there's going to be some prime spots. And I think Arch mentioned it. I don't have to repeat it, but those are all quality spots as well as one other spot that we've talked about in the last couple of weeks is on the other side of AJ Turrell. Who's going to be that guy who can solidify that spot? I think the guy that Arch was alluding to was a guy like Patrick Sartain who can go on the other side. And you've got two shut down corners in a league where you know it's going to be a past frenzied league, not just even in NFC South, but in the entire league, teams have two, three receivers. Look at us here in Atlanta. We've got a bunch of receivers. There's not a lot of DBs that can match it. If you can get another guy on the outside of their corner spot too, that bows well as well. Yeah, there's definitely plenty of options with that number four pick. And I think you guys have talked about a couple of scenarios where maybe Atlanta ends up trading back because DJ for Kyle Pitts, four might be a little bit of a stretch for him, but 10 or 11, probably a little bit more in his range. And if Atlanta decides to go quarterback and it doesn't end up being obviously Trevor Lawrence or a Justin Fields, maybe they end up trading back a little bit and you still get a lance out of North Dakota State or something like that. Again, a lot has to happen between now and then to see where these guys actually end up slotting. And again, we could sit here and look up at mock drafts on the internet all day long. It only matters what Terry Fontenot and Arthur Smith and the rest of their staff, how they end up ranking the board. That's the only thing that matters when it comes to the Falcons and who they're actually going to address. And, you know, the other thing we talked about guys, we're not going to dive too much into it here is the salary cap obviously plays a huge part in a lot of this that goes on because we've talked about the draft, but there's a free agency that ends up being a very integral part of building your roster as well. But it's tough to dive into that right now with the salary cap. And if you're one of those X's and O's or excuse me dollars and cents type people, there's plenty of websites out there where you can learn about the salary cap because as Dave talked about before, we got in his podcast, there's one, maybe two guys in an organization that could actually explain how a salary cap works. So if you're Joe Schmoe watching the podcast in your living room and you've got the answer, put your application in for a team because if you could figure out the salary cap, right? That's like us. It's three of us over here and Arthur's the only guy who knows about salary cap. So that's pretty much how it goes in organizations too. I bluffed my, I bluffed my way through it. You guys bought it. Hey guys, I wanted to, let's, let's kind of end this with this thought because we've talked a lot about both of these guys that have gotten their career started with the Atlanta Falcons. Arthur Smith is the head coach, Terry Fontanose. Now the general manager, a lot of work ahead for those two guys, probably going to have very little sleep the next few weeks, probably going to have a whole lot of phone calls and text messages, whether it's congratulatory or calling people to build out their staffs. But let's just take a look ahead. And again, it's hard to really assess what it's going to look like because these guys have not been leaders of an organization, whether it's a general manager or a head coach, but DJ, after what you saw today, how excited are you for the future? And, and I don't want to sit here and say, did they get the right guys? Because that's such a cliche question. But what stuck out to you that gives you promise for 2021 for the Atlanta franchise? Right. This is something I learned a long time ago and it's going to help me become the, the man I am today. And it's first impressions are everything. And today was my first impression of Arthur Smith and Terry Fontenot. And I was truly impressed by the way they handled themselves in their press conference. Now it could have gone a number of ways. Of course there's going to be a lot of time to tell about how these guys really work out in this organization. But when I look at Terry Fontenot, there are two things that really stuck out to me.  And you think about, if you watch their press conference, you saw him bring his kids on there. You saw, he talked about how excited they were. He talked about now there's going to be a bunch of Louisiana Falcons fans now. And then the other part that really impressed me was at the end, before he started, you know, getting questions, he did something he didn't have to do. He made an acknowledgement of a guy that we all know very well and Vaughn McClure and went on a, a, you know, a nice little soliloquy of, you know, five, 10 minutes about what Vaughn meant to him, but he meant to a lot of people. And I thought that was, that was paramount. That's him. Like that's something you can't fake. That's something that, you know, he's in a major position. And this was all about him.  You talked about it. He, he choked up talking about Mickey Loomis. Like that kind of stuff tells you what kind of person a guy is. And then listen to Arthur Smith being in the position. And the one thing that I heard coming in, I'm going to keep it all the way, keep 100. Everybody talking about, ah, I don't know how he's going to be with the media. He really hasn't had much training. I thought he was awesome. I thought he handled himself well. I thought he answered the questions that were presented in front of him and then shy away from anything. So first impression for me or everything, and both those guys kicked it straight through the uprights and maybe the first touchdown on the year for me, for sure. All right. Hopefully the first of many to come for the Atlanta Falcons. Arch, you're going to be speaking of touchdowns. You'll be calling many of the touchdowns as the analysts for the Atlanta Falcons radio network. And so again, after you saw Fontano and Arthur Smith today, just give us your kind of overall impressions in your excitement level now that you know the top two guys outside of Arthur Blank and Rich McKay that are going to be running this organization from a football perspective moving forward. Yeah, I'm ecstatic guys. I really was impressed with both guys. I would concur with shock. I thought both guys handle them extremely well. Remember, this was a preface conference for the coach at one o'clock. It was a press conference for the GM at three o'clock.  Both of them said the same things. And not that somebody was beaten on their head or handed them a script. This was all stuff that they believe in together. And that's pretty cool that you got two guys that are in lockstep with one another that well, the question I have guys is for coach for, for cherry font. No is how does he get rid of all the same stuff? You know how you can't get any of the same stuff through Atlanta, right? He's even made reference that it says saints on his credit card. He says he's got to get his credit card back. So you talk about it and it's funny and we could go tongue in cheek with it. But think about it. This is a guy grew up in Lake Charles. His wife grew up in Lake Charles. They got married in New Orleans. He's been with the same since he was 22. He's 40 now. His kids don't know anything, but going to Saint games and, and cheering on Drew Brees. And I thought it was really cool right near the end of the press conference.  And one of the guys on the press corps said, Hey, Kate, who's your, who's your favorite football player? He said Julio Jones. I said, wow, you talk about being buttoned up and ready to go. Now the family is dialed in. They're ready. I mean, come on with a kid that age, how easy would it have been to say Drew Brees or Michael Thomas or something like that? So yes, it's, it's almost. For goodness sake. He didn't even have a, he didn't even have a red tie. He said he had to go get a red tie and literally right before the press conference. That tells you how long he's been with the station and not having any red and black. Two humble guys, two humble guys that both mentioned leadership by servitude and that's who they are. That's who they are. That's who they are. They kind of came up, kind of grew up through the ranks. Some of it was a little quicker. Maybe for Arthur Smith and it was for, for Terry font. No, but they both worked their way up. And you, you can't say enough about Arthur Smith being retained by three different regimes in Tennessee. Kept him as a coach. That doesn't happen. You might be kept over one time to be carried over three different times. And really it was four times because malarkey and, and, uh, wasn't that kind of shared the duties. After a month check. So it was actually four coaches that kept him in Tennessee. Tells you a little bit about who he is. And then the giving, being given the offensive coordinator duties, his final two seasons and improving on what they were doing in 18. Oh yeah. By the way, that guy in 18, he's now the head coach in Green Bay. That worked out pretty good. Right. And this guy blew his numbers out of the waters. The offensive coordinator. And that guy tried to steal him to the floor. He tried to steal him. Pretty good. Pretty good. Well here, here, I'll leave, I'll leave it with this. My, my impressions are really quick. I thought our Smith, he looked like a football coach. Sometimes you get a head coach that gets up in there and they steal the press conference because they're so, they're so clean and buttoned up and they got the $3,000 suit on. But when I saw Arthur Smith, I was like, I don't know, I don't know. I want this guy to know X's and O's better than everybody else in the league. I want him to be able to make adjustments better than anybody else in the league. I don't need to have that guy know how to do the sweetest tie knot on his tie. Okay. Terry font. No, on the other hand, he was really polished. I was impressed by his professionalism. I was impressed by his plan, even though he said that being a general manager, I was impressed by his professionalism. I was impressed by his professionalism. I was impressed by his professionalism. Even though he said that being a general manager wasn't his plan. So I feel like the Falcons are in really good hands with these two. Even though these two don't know each other that well, I'm sure they've been on the phone a whole lot these last few days trying to get to know each other better because they're going to have to work hand in hand over the next, hopefully a long number of years, let's say five, 10, 15 years because we know sometimes the NFL stands for not. All right. That's going to wrap it up on behalf of DJ Shockley and Dave Archer. Just bring in the thunder as always. And DJ says he keeps it 100. Archer, you keep it 100 as well. We thank you for joining us on the Falcons Audible presented by AT&T, a special edition to talk about the Atlanta Falcons new head coach and general manager. And guess what? There might be some more breaking news coming down the road and you'll get us three back on and we'll be here to talk about it. Thanks so much for joining us everybody. Once again, on behalf of DJ and Arch, I'm Derek Rackley saying so long, this is the Falcons Audible presented by AT&T. You're listening to Falcons Audible presented by AT&T.


I'm Ryan Poole, Storm Center Operations Manager at Georgia Power. Our teams around the state are ready to meet customers needs during these frigid temperatures this week. We are monitoring the weather very closely and are preparing our teams to flex line crews around the state as needed. From a weather perspective, we anticipate the threats to be windy conditions and cold conditions on Friday, cold conditions on Saturday and Sunday. Not only are teams prepared to meet any issues on our power lines and other equipment, our generation crews and generation plants, just like the one behind me here at Plant McDonough, are ready to respond to what we know will be increased demand with a low temperatures. We anticipate weather like this and have plans in place to protect our equipment from freezing temperatures. We invest in our generation fleet, invest in our grid to ensure that it can maintain reliability for our customers even when the temperatures get in the single digits. On behalf of our team at Georgia Power, please stay safe and take care during this holiday season.


Thank you for this opportunity to speak here in this amazing venue. I'm going to talk about some of the work we do in my research group on yeast. So we do engineering of all sorts of different scales on synthetic biology systems in yeast and probably about two-thirds of the work in my group. I was going to build up to some of our stuff and eventually talk about work on the synthetic genome project with yeast but Jeff is coming after me and he's going to give a much more detailed talk on that so I'll probably concentrate a little more on some of the stuff at the circuit level and maybe at the individual gene level in terms of using modularity as a process and maybe if there's time I'll talk about some other work we've done that answers that sort of answers more some of the questions that were posed yesterday. So the my group generally works on foundational tools a whole variety of different things with the name to try and make engineering of biology more predictable and something a bit more akin to an engineering discipline and we started now moving to apply some of these tools in different areas diverse areas such as materials production and therapeutics production in particular. We kind of see that within nature there is some level of modularity most genes are considered as modules you see operons as modules and these scale as you build up chromosomes built from these various modules and so we can also see comparisons and this is from a Ron Weiss review paper with how modularity allows sort of scaling of complexity in other disciplines and how you can also see that naturally in biology. I'm not a firm believer that these modular rules exist and they're never going to be broken. Biology is very good at finding ways to break rules but I still think that as we do work in synthetic biology and constructed different levels being able to use modularity gives us a lot of power and enables us to really progress in the subject very quickly. So I first started getting involved in synthetic biology in Jim Collins's lab back then at Boston University and what we did was work with modular parts rewiring them into different circuits and you can see here a couple of different circuits we published on one of them being a feed-forward loop network and one of them being a bistable toggle switch which allowed us to look at how noise is it influences in differentiation of cells. Both of the networks you'll see here contain pretty much the same parts and the same modules, promoters that are regulated at different times by different regulators, bacterial regulators in this case that we've moved over into yeast and placed in the yeast genome and by just simply changing the topology of the interactions of these regulators we could get different effects which is kind of a way where just with a very limited number of parts we can get a lot of different diversity in the behavior of yeast and so just to give you run through one of the examples of these projects we wired up these two bacterial repressors against one another in a what is effectively a leaky bistable switch system and this became a way to program in predictably the timed message within the cells of gene expression turning on it after a certain period of time so we called these kind of like slow release genetic timers. So just to talk through this network design here we have two regulated promoters that are producing two bacterial repressors these repressors when expressed will repress the expression of their opposite repressor so that you have either one system dominant one side of the system dominating or the other but because this is a leaky system almost always the tet R side of the system would dominate unless you came in and gave a signal to the cells which was anhydrotetrocycline you could also give doxycycline which blocks tet R from binding on its repressor and will then instead see the system start producing more lack eye so you can see here in our normal system we start the cells they immediately revert to their favorite scenario where tet R dominates the system we add anhydrotetrocycline for a period of time and then we can wash it off the cells that has flipped them into being a lack eye dominated system and then over a period of time due to the the leakiness of some of the promoters here you end up with the system slowly reverting back in this case over about two days worth of continual yeast cell growth so that's many generations of yeast so this is a message that they're passing on to their progeny not by changes in the dna so it's a form of epigenetic memory passing down and so this system very much works like if you had a switch on the wall and you flipped it up and then it it slowly went tick tick tick over a period of time and then went down and we could plug this in to our yeast and see gfp levels change as the population grew we then realized that because the nature of how this time of reset was to do with how much leakiness there was in expression from the promoters and how how well they were controlled if we did point if we did a general mutation of regions of the core of these yeast promoters we could get a whole library of promoters that led to different expression levels but also different leakage of expression and we could then use these different regulated promoters in our system and we could take data from this part library put it in together with the prototype data from the initial version of the network we made and we could use something we could use predictive modeling to kind of work out different reset times from different combinations of modular parts we're putting together so the way we do that is quite straightforward we build our network we do mutation within specific regions of the promoters to get our promoter library we characterize the promoter library for both that output how much they produce when unregulated and their and their kind of their input which is how repressed they are when tet aralakai is overexpressed and this data can then be fed into a predictive model and this was done by Jia Wang in Jim Collins's group and we can then look to see when we have different combinations from these part libraries how long the reset time of these genetic timers would be and these blue lines of the prediction space for these things and the red lines where actually when we went and took a few of these combinations together and then measured the reset times this is what we got some of them nearing almost a week of exponential growth before the reset gets all the way back to normal this is a good example of how modeling can be used in synthetic biology projects especially circuit projects like this we had three promoter nodes in our system here and we had 20 promoters in our each of the promoter libraries that could go in here so we could have produced 8,000 possible networks that actually could be done probably now by PhD students in good groups with good tools but back in the late 2000s when I was doing this work that would have been a hell of a lot of effort by me and Jim's lab with the basic tools that were available so it's really nice then that by having a predictive model we can then use that to define a small number to build rather than have to build very large numbers these networks were only controlling GFP expression so we wanted to show that they could be used for something slightly more interesting and we managed to discuss with Kevin Verstrapen's group who was then at Harvard about an interesting gene flow one which when knocked out of yeast or under expressed in yeast the yeast grow normally they you know they float around within the liquid perfectly fine but when you over express it in cerevisiae yeast you really start to get tight binding of the cells to one another and this is because it's this protein which forms on the outer surface of the yeast cells and acts like velcro getting cells to start linking together we call this process flocculation and so when you get enough cells flocculating within a population they then have a gravity and so they then set them down and you end up coming in in the morning and seeing this pellet at the bottom of your tube and actually if you go to the microscope you often see very very tightly packed yeast cells sometimes even looking like hexagonal when i wanted to get microscope images in the lab sometimes i would have to be fishing these pellets out and then smashing them and breaking them off with pipette tips to be able to get some that i could see under the microscope so this is a really cool natural glue that yeast has so we linked up our timer switches to this system and now the different timers with combinations of different promoters in the key nodes of the of the circuits can now give us sedimentation and pellet formation within our exponentially growing cultures at different predicted times and again going on one here almost a week afterwards so this was a nice way where we could tie in a really interesting phenotype of yeast with genetic timers that we built and we'd use predictive modeling and we could get new kind of phenotypes seen and control of that as well we've been kind of pushing that a little further in my research group inspired by a royal college of art fashion designer who came to our group for a few months with a wacky idea of that it would be great in the future that instead of mass manufacturing of clothes where all the clothes have the same pattern instead you have microbes performing the patterning so that every one of these mass manufactured items of clothes has an individual pattern so she investigated how you could grow bacteria in patterns and we got bacteria to secrete a pigment molecule as this was as they were growing and we even tested this out on fabrics and showed that this was okay to then autoclave and take out of the lab with this patterning but it got us thinking about whether we could ever make patterns with the way yeast grows so yeast normally grows obviously as a single cell organism which then buds off and then another single cell organism grows but in a colony what happens is they all start just growing on each other but yeast also has hidden away in its genome it has the genes that allow it to do kinds of multicellular growth most of these have been have got now mutations within them that silence this because the lab yeast we use we don't want to see that happening but you can resurrect these mutations and start getting things like filamentous growth, pseudo hyphal growth out of yeast things that are seen in natural yeast and filamentous fungi so there is a poster I think outside here where someone else is looking into this is the person who's posted yeah I haven't had a chance to talk to you yet but it's super cool stuff so we were we then wired up these some of these genes my phd student was very happy that one of the key genes involved is the gene phd1 so he gets his phd straight away with the mutation there and then so now we have using the same promoters that we used in the previous network we can link up to control of flow 8 phd1 and also another one cdk8 and so now we can have extra extracellular control of filamentous growth so that these cells instead of growing and budding off from one another when we hit them with specific induces they now grow like this where they don't leave one another they grow more elongated with branches coming off so they're starting to look a little like a branch fractal but one that we don't have much control over so just to show you how they grow this is kind of like filamentous growth where we've induced this and we managed to do this in haploid strains which was the first time people had had done that because it's normally only seen in diploid strains and you can see the branches come the filamentous growth coming off and occasionally branches coming off that as well and we can put the whole system now wire it up so it's linked to our genetic timer switches so over a period of hours and days you see filamentous growth initially and then as the time has started to release the system you'll now start seeing them start forming colonies as normal so they start filling up in the gaps between the branches. My student George is finishing off work on this we currently have unregulated branching which is which is great for making filaments but actually being able to control the pattern really requires us to be able to control the branching rate and particularly when and where how often you get a branch forming so what currently happens is a cell will divide off and make the neck cell and this will make the filament so this is we call it in yeast the mother cell and then the daughter cell and what we're trying to achieve now is differential regulation between the mothers and the daughters so that the mother cells then enter a period after they've produced a daughter cell where over a certain amount of time or a certain amount of gene expression they don't produce another daughter cell so that prevents them making another branch but then after maybe several hours or days we could see that signal come back down again to allow it to then produce another daughter and that would then produce another branch into the pattern here so to get that we're working very hard on regulated promoters that are regulated so that they're only expressed in the mother cell and not the daughter cell and those can be tied up to senescence genes we have some initial work there that I'm not going to show here that that seems to show that that is working as well that kind of piggybacks on some of the work my group did earlier on a few years ago where we looked into designing promoters from the bottom up because what we need in that scenario is we need a promoter where we know we can have repressible externally repressible expression but we also needed that the regulation turns on in mother cells but not daughter cells that's a complex conundrum for us because we have to take regulation that is found naturally in yeast in upstream activating sequences but we need a core promoter that can be repressed so we're starting to have to make hybrid promoters where we're bringing in different regions from different promoters putting them together so that our logic can be like we want it to be so we started doing work towards this end on a different set of promoters a few years ago and this was published way back in 2012 was one of the first papers from my group when we first got going five years ago and this was we looked at could we kind of start with a very very boring promoter that worked as a promoter so that genes would be transcribed in yeast and then could we begin layering on our own regulation on that in a more predictable way as opposed to taking initially well regulated promoters and trying to work with those where they have a lot of things written into their sequence which define when and where they're switched on so we did possibly the world's most boring bioinformatics screen which was we looked at thousands and thousands of gene expression data banks that were available online to look for mRNAs that almost never changed their gene expression in lots and different conditions because we wanted something that was just on and always on this gave us a sort of a top 10 list here and one of them we settled on was the profile in one promoter PFI1 this produces a cytoskeletal protein quite a small one and what we really liked about this promoter apart from being very very constitutive in that its expression levels don't change much is that it's also very short and very simple it simply has a poly T-track which is bound nearby by red red one protein and then it seems to leave a core promoter region here which RNA polymerase can keep binding to and firing to just keep doing gene expression of whatever you put downstream of this gene and so we tested that in a variety of different conditions here and we always saw very close constitutive gene expression at different times in exponential growth of yeast and in different conditions as well and then we did this trick again that we've done back in the the previous work in 2008 where we mutated bases just in a very specific region in the core region of this promoter and we showed that by mutating all these bases you could select out a whole library of promoters that the slight changes in the base sequences there now lead to the promoters having different strengths but they're all very constitutive that also showed us as well as giving us a library to do lots of different levels of gene expression in yeast it also showed us that there were regions within that core promoter where you could change the base sequence you would still get output so that means we now have a kind of an area within the promoter where we can start adding in our modules so a simple one we did first was to add in the recognition sites for the bacterial TET repressor so that this now switches from being a constitutive promoter to being a regulated promoter that is usually repressed in the presence of TET repressor and then as you add in increasing amounts of anhydrotetracycline you can switch this on so this allows this now to become a regulated promoter at the time back in 2012 the the new cool invogue thing or 2011 actually was tal effectors and we showed that tal effectors which are very modular dna binding proteins where you can swap out domains of the protein so that they target any pretty much any different dna base sequence we could then get them to target the different sections of our promoters and they would be what we call orthogonal which is that one repressor would specifically bind one promoter and switch that off whereas the presence of another repressor would not interact with that promoter and so that you would have a very defined relationship so now we're able to scale this so that we've gone from one boring pfy1 promoter to now having multiple regulated promoters with multiple regulators for each one dna 2.0 offered to analyze our mutation sequence from this library they went through this sequence and they have an algorithm that looks to add all the base sequences within a mutation library and what the output of that promoter was and then tells you the relative importance of every base as well as they can from the very small data set we provide them and what was quite cool here is this led them to initially say to us okay we we think based on this sequence analysis we could design you a strong promoter from this relationship here and this is the one they designed pretty strong not the strongest but pretty impressive that they get to second best just with their first gas but this also gave us much more insight into the bases that are available within this region to mutate which ones we need to keep as a certain base if we want to maintain strong gene expression and which we can tolerate all sorts of mutations with and so then for designing all of our further regulation we found that there was a specific region where we could change a number of different bases as long as we kept this t and this a the same and we could generate all sorts of different targets within that region which could all then be individually targeted by different telefactors but the gene expression that would come out from the promoters would be exactly the same in every case so you have a whole whole load of different regulated promoters but they behave almost exactly the same both when activated and when repressed and so we haven't yet published this work we're kind of wrapping it up at the moment but this is how the orthogonality scaled this was the six by six matrix bends taken this up now to a 10 by 10 matrix where you can see all the different telefactor-based repressors match and repress down only at the specific promoter pair that they're supposed to be with so this gives us a whole lot of different tools to do the rewiring here just to cover we've also done other promoter libraries here this is constitutive promoter library this one's based on the very strong adh1 promoter with a whole range of mutations and this was done in a slightly different way because this is a much longer promoter region where we used highly mutagenic PCR to introduce about 10 errors into the promoters and this again leads to a different level of expression what was kind of cool about this was this was done in the same pot as doing DNA assembly to make the plasmid at the first time and this was part of work and we've done lots of work in my group and with Jeff Baldwin at Imperial College on different DNA assembly methods and this showed that in using a certain method you could in the process of constructing your plasmid you could say I would like this part within the plasmid to appear as a library of mutation in the same go as you're doing the whole experiment and the assembly so on the same day that you assemble your plasmids you can also be defining that one position within that network or within that pathway is mutated so DNA assembly is something we've like I said we've done a variety of different papers and reviews on and I just want to now move on to talking about some of the success we've been having since we moved to a golden gate-based assembly we aligned ourselves with John Duba's lab at UC Berkeley who developed this fantastic MoClo yeast toolkit which allows you to take a whole load of different modular parts assemble them together first to make kind of a gene system so a promoter an open reading frame a terminator and then you can take multiples of those and then bring them together to make pathways or to make signaling pathways or just multi-gene systems and the kit that you can go out and get now an ad gene if you're a yeast group and you want to get using this contains a whole variety of different parts there's about a 96 well plate is what you get in our own lab and in John's lab the these collections have now ballooned to many hundreds of parts and actually John sent me this slide a few a couple months ago looking at when his lab switched to everyone on the yeast projects using this specific framework of modular MoClo based assembly the massive increase in the productivity of the lab in terms of how many plasmids they're they're all making and I certainly think it's been the same in our lab since we moved to that about a year and a half ago so we've been mostly using that for pathway engineering and we heard plenty about pathway engineering yesterday without having tons and tons of mass specs like they do up at Manchester we've instead been looking at pathways it's very easy for us to to see a good output for so Jeff's group have been helpful in sending us over some pigment pathways we've also been working on selectable pathways such as the production of the antibiotic penicillin and selectable growth pathways a commonly used one in yeast research is introducing the genes that allow yeast to grow on xylose a lignocellulosic sugar that is it doesn't normally grow on penicillin is probably the one of most interest and it's one that we've really enjoyed working on and we're just wrapping up the work now as a paper and so this is how penicillin is made firstly a gene condenses two amino acids and an unnatural amino acid into a molecule called ACV which is very quickly then modified to become isopenicillin in the peroxazone of the filamentous fungi penicillin this is then modified to make PENG which is then secreted out of the cells and this goes out into that external media outside the penicillin chrysodium cells so we decided to put these pathway libraries these guys all on a on a plasmid to add to yeast this first gene here the ACVS is a type of gene called a non ribosomal peptide which is a assembly line enzyme which is effectively one protein but it's actually encodes multiple different enzyme subunits within it catalyzing lots of reactions as it goes along so you can imagine this as as one sort of multi modular enzyme and as such it's very big it's almost bigger than any gene naturally found in the yeast genome so it's a bit of a pain to clone because 12 kb you know PCR is kind of off the table unless you can guarantee really good accuracies but we managed to clone this and have it under a regulated system into the genome of our cells and so that goes into one of the chromosomes the rest of our pathway goes on plasmids and we use these modular dna assembly kits to put in some different promoters different versions of the open reading frames that encode the enzymes adding on tags and different three prime utr or terminator sequences because that can also affect gene expression in yeast and when we put all these together combine them first into their individual gene plasmids and then assemble them into a final yeast pathway plasmid we can start getting ourselves producing penicillin so firstly this is the pure chemical standard and the mass spec trace shows that that big gene on the enzyme is working so we get the non-riversal peptide synthetase working in yeast and it produces the first part of the system which is ACV and then the rest of the pathway then allows us to get PENG production and what's really great about this actually is that the PENG is highest in the external media because as well SRVCI is secreting this out naturally so we find much more of the penicillin G in our growth media of our yeast than in the actual cells themselves which so that makes life very easily and what was great about the Moclo yeast toolkit in this system was that the yeast toolkit included localization tags to send certain enzymes into compartments and one of those was the paroxysome targeting tags so for example it we when we got the kit from john's group we immediately, Ali immediately stuck fluorescent reporters with these sequence tags on the end and turned on the gene PX11 which causes overexpression of paroxysomes in the cells and we could see all of these expression now of these proteins these enzymes being sent into the paroxysomes and many more large paroxysomes in our cells and that really helped us boost the yields in our pathway and especially boost the yields we were getting in the extracellular media as well because so much more of the reactions are occurring now in big paroxysomes that are fusing and leaving our yeast. We've also slightly improved in the last few months this Moclo CRISPR system, Moclo system to include CRISPR so now we have a very fast protocol in the group for putting in a variety of different things into the cell at the same time and at the same time making changes on the genome and what happens here is this is so efficient now that you do not need any selectable markers on the things that you're integrating in because in yeast the CRISPR is cutting the genome until you fix the sequence there so that sequence doesn't exist and all the cells that can't repair that sequence are going to die and you then provide in donor sequence maybe the gene that you want to add in or the mutation you want to put in to the to the yeast chromosome and that will repair in very efficiently because yeast is very good at homologous recombination and so then you will end up with all your changes. So Will first go trying this whole protocol. He had greater than 90% success rate with no selectable markers doing simultaneous two deletions and two insertions into the genome. So this is now scaling up and so the sort of things that people in my group, Will is just a first-year PhD student. He's now doing things like looking at signaling pathways, looking at doing multiple knockouts of different components at the same time as putting in refactored components now with a whole variety of different promoters to change the gene expression of the components in a singling pathway and so in in a one-shot experiment here as an example he knocked out four genes on the genome and added two more genes back in but under a library of different promoters and was able to pick out colonies that then maintain the signaling systems so they still respond to Mf alpha this mating factor so that increased mating factor leads to increased expression from the reporter gene not quite the same as wild type it's a bit more leaky but if you see if you haven't added these genes back in the whole thing is constitutively on and so this is pretty impressive because this is six different changes going on in just a in a one-pot reaction in the cell. So the last thing I was going to talk about on yeast was the modular least yeast genome synthetic yeast but I'm gonna let Jeff give you the whole introduction here but I just want to point out that the reason why I call it the modular yeast genome and why I'm so interested in working on it is because it has this recombination system written into the chromosomes that I'll go into in a second. Our lab is involved in building one of the chromosomes chromosome 11 with the kind of scary number of six six six killer base pairs in size and I'm happy to report that we've got all of our DNA it gets synthesized for us by genot and gets sent like this it's very boring it's just a FedEx package that costs about the same as a small car and then Ben in my group with the assistance of Maureen has been putting this together and 76 percent of the synthetic DNA is now integrated into our chromosome so we're nearly done and you can see our progress on our resource web page along with the progress of some of the other chromosomes in the project but why I'm calling it a modular genome is because the design that Jeff and others came up with was to place a recombination site locks piece in landmarked throughout all of the chromosomes immediately downstream of all non-essential genes and other significant landmarks this is a recombination site where if you provide a recombinase Cree that recognizes this site and you add estradiol which is induces it to go into the nucleus it will bind together two of these sites link them together in a loop and catalyze a rearrangement of the DNA leading to either translocations so that one part of one chromosome shifts to go to another part of another chromosome inversions so that a region of the genome flips over so it's now pointing in the other direction and also deletions that's a very important one the genes could be lost and potentially as well insertions if you provide new DNA at the same time that's within this format and so this is from one of Jeff's papers this shows that the majority of the time if you have synthetic chromosomes in your cell and you induce this system most cells are going to die and so the reason for that is quite obvious because they're deleting or rearranging essential genes so that the new versions of the chromosomes that arise from this recombination are not viable so you get lethalities but what's very cool is that many of these colonies here there are survivors right so these are cells that have maybe slightly rearrange their chromosomes or even dramatically rearrange their chromosomes but they're still viable so the modules of the individual genes have changed in both in terms of that content some of them might have disappeared some of them might be present in multiple copies instead and the arrangement of them have all shuffled around but the cells still grow and so I'm very excited in this because gene expression has been shown by others to be significantly affected by the location you are in in the genome and things like the secondary metabolite pathways like the penicillin pathway we're working on and others are often found to cluster in plants and fungi you would have seen that from an Osborne's talk earlier this week and here's an example of a metabolic pathway that's clustered through evolution in S. cerevisiae the dial cluster and so the modular genome of synthetic yeast allows us to to look at this in a way and try and understand how a genome could be laid out to do things like optimized biosynthesis and then eventually one day I would love the challenge of of having a full set of parts and thinking from bottom up not copying what nature does how would you put these various modules together to make a viable genome from scratch to give an example of some of the effects of genomic positioning with tools that were sent over from Jeff's group we put in a violation pathway in different positions around a partially synthetic chromosome putting it in the middle of the chromosome putting it up close to the telomeres spreading it out so they're at either end of the chromosome close to the telomeres and the production of this pathway if it working perfectly well violation pathway should produce some very dark purple pigmented colonies and what you can see is certain versions of these constructs do that very well others the whole system doesn't work possibly because of local silencing close to the telomeres and other ones are a bit mixed and so we even occasionally saw colonies that were half white and half pink because some sort of silencing effect was going on within that colony where we're going with this kind of work is that we think that the scramble plus the heterologous pathways is is a good interest so we could put our carotenoid pathway pen pathways isle pathways in cells that have partially synthetic chromosome regions or even full synthetic chromosomes we can induce this scrambling system and what that does is that creates a whole diversity of different yeast strains that your metabolic pathway is operating with and some of those may be more favorable you can even rather than construct your whole pathway you could provide your genes as just units that are ready to be scrambled in and we did that with the xylos pathway which is two to three genes best if there's three but it can survive with two and we were able to do this scrambling and pick out a strain which actually started to grow on xylos media because it must have inserted these genes in the genome so where I think this is quite useful is in the metabolic engineering world linking that in with synthetic biology the traditional method is quite to get high yields is quite multi-step you would initially design and clone a version of your pathway put that in your cells test it and then you'll go back and start swapping in alternative genes different promoters of different strengths to try and optimize the expression and flux of the pathway you'll then probably take your best winners out of that take them out and now move them into lots of different strain backgrounds so you might do lots of different strains that people have worked with before that show good yields in industry we might do put them into knockout collections because it may be that certain genes when knocked out will help improve your pathway and then finally probably the one that in industry really matters is optimizing the growth conditions for scale up and so all these tools the modular DNA assembly with promoter libraries part libraries plus the synthetic yeast can really narrow this down so that this whole process can all be done so you can have one pot assembly of many thousands of versions of a pathway and you can have one pot strain evolution so that your your best pathways here can then be just scrambled so that the all of these different strains are created in the same pot and you can then look for those as long as you've got a good selection system and you can go straight up to optimizing conditions to get your better yield and this is part of a project that I had fun I've got funding for at the moment where I tried very hard to get a really good acronym for my project so this is it automatic chromosome rearrangement for optimizing novel yeast metabolism I was very glad the funders saw that as a good acronym and a lot of these methods that we're developing we're going to be teaching next summer at a synthetic genome engineering summer school that I encourage you to send your students or yourselves ask your supervisors to go for it'll be heavily discounted so it'll be very good value five-day residential summer school up in Edinburgh looking at things like crisper mediated integration synthetic yeast chromosomes scrambling evolution and it's going to then lead on to a brilliant two-day conference on synthetic genomes and synthetic yeast which Jeff will give a mention of and a final thing I would like to shout out is that there is still time if you have researched that Patrick Tai up in Edinburgh and myself are asking for people to send in European based research to a special issue of ACS synthetic biology this was originally with a deadline at December but they've extended that now so you can get it in by about mid-january so let me know if you're interested in sending in any research so I'll just wrap up now because I guess I'm almost out of time and thank my group this is most of us proving that the the density of rock is significantly more than the density of humans so whatever we do we can't unbalance this rock up here on the top of a mountain in Wales and I particularly want to highlight the people whose work I spoke on Ben who worked on the telefactor and the promoter library stuff and who's also now leading the work on the synthetic yeast chromosome George who's been working on the patent formation with the yeast colony growth from filamentous form and I'd also like to particularly mention Ali who's doing all the work on the penicillin pathway and the collaborators here and especially I'd like to thank Jeff a great collaborator as well and my funders so thank you we can take a couple quick questions for Tom and then we'll take a five minute break before we have Jeff's talk right so I was interested by what you said about the the position the genes that encode the secondary metabolism pathways they often cluster in the sub-telomeric area and this is as you mentioned is known to to correspond to science regions in in general however it has been shown at least in the case of low-level irradiation that some stresses will rip off the telomeres from the nuclear anvil and will end up in reducing the silencing that is in overexpressing the sub-telomeric and telomeric genes so that in some sense could it be first of all could it be that chemical stress or has it been shown that chemical stress will induce this response by inducing the pathways of secondary metabolites like the ones you mentioned is it known and second of all is it possible that or do your data speak to this phenomenon you have shown that you have put your pathway genes at different positions could it be interpreted could the results be interpreted in in a way that says that they are induced they are they are silenced near the telomeres and that they are induced by any type of stress okay so it's a really good question I'd not heard that hypothesis before so there's I speak with Anne Osborne about this quite a bit there are generally a couple of hypotheses why you have clustering of secondary metabolite pathways in things like the plant genomes and the filamentous fungal genomes and so one of them is deemed evolution that by having these genes in the sub-telomeric regions where you're more likely to get crossovers between chromosomes it leads to quicker evolution of new secondary metabolites another one is co-regulation it's obviously you don't want the secondary metabolite to be continually expressed because they're often very stressful for the cells so it'd be nice to have them in an area where it's easy to co-regulate and silence the whole one I think your hypothesis that adding a stress that then turns that on is a is a very good idea as well that would I could also see that being a benefit the problem is I don't think anyone has studied that because these pathways are where they're naturally seen is in hard to grow hard hard to work with filamentous fungi and with plants as well so those kind of experiments very difficult so one of the main reasons we're doing this project this acronym project is to put in a selectable secondary metabolite that the cell might use to defend itself in a way which is what some of these secondary metabolites in the plant systems are so the production of an antibiotic so that it can then compete with bacteria that are in the growth media and so that we have a system then with in yeast which is very tractable where we can then start to ask a whole load of these questions to see whether these things you know are good can we have the whole thing repressed in terms of its expression and then induce a stress and we will see more survival that's something we could test it's a really nice idea so just as a follow-up I would like to to make it clear it's a fact that low-level irrigation rips off the and overexpresses the subterlomeric genes this is fact the hypothesis is that it could happen with other types of stresses and no idea for some reason would it be possible to use the flocculation for forming two-dimensional patterns instead of the pseudo-hyphen growth I think so I think when we started that project we looked at a variety of different things and we just saw that the pseudo-hyphen growth was very kind of close to where we wanted to get to anyway which was these branch fractals so we stuck with that but yeah flocculation is a fantastically interesting phenotype as well did you try growing them on like a plate or expressing effect with the flocculating strains that I showed at the beginning the flow one strains yeah they would form pretty horrible colonies unless I was repressing the the flow one gene and then but most of the experiments we did way back when we were doing that flow one over expression was mostly focused on we would keep the colony not we would turn off flow one in colony growth and then switch it on specifically for the exponential growth phase so that we could then get those pictures of the different tubes with the palette forming over time Tom so I guess another question relates to the growth-based patterning I was wondering is there is there a particular advantage to doing kind of growth-based as opposed to kind of cell patterning techniques you know essentially printing cell type printing patterns that people have done like Zezart and just a lot of UCS stuff for example you know where you have your pattern you just print the cells even different types of cells and things like this is there well so one of the justifications for doing this project and actually it's kind of cool because the funding agency the labor whom trust who have this nice charity in the UK they actually encourage you to submit projects that you know you can't have a good justification for like in terms of commercial they say you know their remit is to fill the gaps between other research councils for the interesting cross-disciplinary things but we did we do think that in 2D growth a growing branched fractal is one of the highest surface areas and a lot of what you need for you know testing out things in metabolic engineering might require good surface areas so good secretion as opposed to if you grow the cells as a single 2D colony they're just going to grow on top of each other now if you were to print the cells in a 2D branch fractal we're still going to want them to keep growing because you know all of these pathways run best when the cells are growing pretty well right so they will just start growing and they'll start growing on top of each other filling in the gaps right and so there will be dead zones there will be areas in the middle where the existing cells are no longer exchanging nutrients because they're surrounded by other cells so they don't have access to the full surface area there's a detail about what you said quite quickly about the peroxisomes that would deliver a penicillin directly at the cell membrane so I think in yeast it's unconventional that peroxisomes fuse directly at the cell membrane so is it that you are meaning it as an indirect path yeah so yeah I did miss I said that wrong if I go back to the pathway actually here they do they don't go at like the peroxisome doesn't fuse and the whole thing empties out it diffuses from the peroxisome out into the cell but the overexpression of the peroxisomes led to much higher levels that maybe because there's much more catalysis going on because we have bigger peroxisomes or maybe because now there's more peroxisomes closer to the membrane so it's coming out a lot more but the ratio between product that was secreted versus not secreted went up when we increased the peroxisome so that means that there is a mechanism to transport the penicillin from the cytoplasm to the outside yeah is this part of the machinery that you know snorkeling that's something that occurs in yeast yeah okay question to you or Francois you think that this is a peroxisome merge with the membrane but to induce the formation of filamentous I'm not sure that those two things are related the these these don't form filaments in this project this is a bit different having said that the the big penicillin producer p-chrysogium is a filamentous fungi and it grows as filaments so that it can have good surface area because the genes that yes that induce are involved in the lipid formation okay I mean the question question mark no are the genes that induce filamentous um well I'll have to go and look that one up I don't know if anyone here knows what the pex 11 is involved in lipids


What's up guys, Isaac here and I'm chilling down the car in the morning as usual and It's meant to be a super hot day today. So I'm gonna be 42 degrees Celsius, which is I don't know how much that is in Phony American in Peru units you guys you change over to the real metric system Catch up to the rest of all guys. I'm sorry Anyway, I want to make this quick video Discussing why you should spend the money on books and generally on anything that's Relate to yourself improvement well, I used to be someone who used to allegedly pirate ebooks and stuff like that allegedly and I Did this because you know, I didn't want to pay for my books.  Make kick-ass Torahs recipes Now what this happens is this kind of Makes you It helps you create the habit of hoarding where you just downloading heaps of things downloading hundreds of books that you never read Whereas if you actually buy your books, whether it be from the bookshop the hard copy from iBooks Kindle or audible all your books You will have a bigger incentive to read it because you want to feel like an asshole for spending money or something I'm not using it So you're more likely to actually read the book and get the benefits from it And this is the same with programs that you might pirate or line Spend money on it because you need to get a habit of investing in yourself You're gonna have it off spending money For yourself investment because you have to understand that In the grand scheme of things 30 dollars here and there for a book Isn't really anything Especially when that book can have that one or two good ideas that changes your life So this is what people get fucked up. They think that The book has to have hundreds of ideas that are gonna change your life when reality when you read a book You especially read lots of books. You start realizing that a lot of books have a lot of filler material They have things that don't really apply to you. They have Things that you don't really need However, they might have the one or two good ideas that are life-changing and what's $20 for that?  Peace like subscribe and all that good stuff. I'll be seeing you tomorrow


Cluster analysis is a form of exploratory data analysis. Sometimes it's known as data mining when you're using large data set. And the idea is to identify relationships in those large data sets. Cluster analysis can either be supervised or unsupervised learning. What we're going to do in this course is known as unsupervised in that we are just going to let the computer try to figure things out as opposed to giving the computer examples and having the computer compare the new data with examples which is known as a form of supervised learning. And the form we're going to use is called K-means clustering. Cluster analysis is really just gathering a bunch of objects and separating them into groups of similar objects by exploring these groups and determining how they're similar and how they're different. You can learn a lot about that big pile of data and hopefully lead to insights that will help you make better decisions. All K-means is just the number of groups, number of clusters that we're interested in. And we're going to put the points in the data set, the points in space into these K groups. And we'll define these groups by a center point. Remember geometry centroids? The group's centroid is its mean location and the distance of individuals in the group from that centroid tells us something we can use. And if we can minimize the distance from individuals to the center of a group, then we can know that the people are in the best group and give us the optimum result. I'm going to use an example I got from a great book, Data Smart, by John Foreman. And it's about a middle school dance. I think we can all remember those dances in which kids filed into the gymnasium and then separated into groups, usually boys on one side, girls on one side, and the counselors and the monitors and the teachers on the other side. So that's a simplistic model, but we can use it to help us learn about K-means cluster analysis. Remember we're going to let the computer do this analysis for us, and the computer has to start somewhere. So it starts out by putting three dots into the gym because we want to do a K equal 3, K-means analysis. And it just randomly places the dots which represent the centroid of the groups. Now this doesn't look very good. If we get the computer to add lines of equal distance so that any point between the centroid is equal distance, you'll see that we've got these three odd shaped areas, a red area up here that has a bunch of people in it and some not in it. A area in the middle that's not colored that's got most of the people in it. And then a green area down here in the lower left which has just the foot of this one person in it. So obviously this is not the optimum location of those centroids. So the computer tries again, it randomly moves the centroid around and then solves some basic geometric equations trying to optimize the distance from the individuals that we have here, the students from these centroids. And this is a little better, but it's still not necessarily the optimum location of those centroids. And we humans would know right away that we should put our centroids in the center of the groups, but it takes the computer a while to get there using optimization. And it will eventually, if we give it the right information, figure out where to put the centroid so that people are optimally grouped. And of course once we have them grouped we can start comparing the characteristics of the people in those groups to learn things. For example, the people in the green group turn out to be all female students. The people in the red group turn out to be all male students. And the people down here in the bottom of the wall of the gym are the teachers, which is the basic solution we probably would have guessed at to begin. But we can use K-means clustering, the technique to analyze large volumes of data and help us make good business decisions. We'll talk about that more in the next video.


Hello everyone and welcome to another video. So I uploaded a video yesterday with a guy who refused to get off the topic of backyard eggs with it being closer to the end of the day and debating, being threatened to be stabbed, dealing with people who only wanted to discuss methane or oysters, oh and plus having a shouting match with two farmers. You haven't got a clue.  I wasn't really much in the mood to play nice. What about the bacon you eat with them? My eggs. The bacon you eat with them? Eggs. Now yeah of course I could have answered the man's backyard egg question in more depth for which I have an elaborate response for but it seems insane to me to talk about the ethics of keeping hens in your backyard and eating an egg they drop out every now and then when you're eating the chopped up body parts of cows, pigs, lambs, fish and the murdered bodies of other chickens. I mean this is like discussing the ethical consequences of swearing with someone who shoots people in the face for sport but in case anyone was interested in how you could respond here are some of the ethical implications of backyard hens. People who eat backyard eggs still view eggs as food. Eggs are the furthest thing from food.   Yes in a hen it all comes out the same hole. Not sure why you'd want to eat anything that comes out of that hole but whatever.  It's disgusting but technically not immoral. It's only when these bodily secretions become a commodity that we begin to have a problem. Now because a backyard egg eater still views eggs as food they're almost always eating eggs at restaurants, outside their home, at a friend's house or in packaged products in their house which contain eggs as ingredients. These products will almost always contain conventional eggs from a caged or free-range farm where the conditions are appalling and are definitely not consistent with the backyard hen owner's happy hens happy eggs delusion. One thing people seem to miss is that the backyard egg industry is still part of the same industry as conventional eggs. Yeah they're in my backyard they're happy. You supported an industry that macerates male chicks. Just like in my conversation with this man when you pay a breeder to give you layer chicks to lay your eggs you are supporting the horrible practice of breeding chickens. If you haven't seen my documentary violated I will briefly rehash what breeding egg layer hens entails. Egg layer chicks come from parent breeders who are kept in specific sheds for breeding. The most common practice for breeding parent birds is the introduction of roosters to the barn. There is typically one male to every 10 breeder hens placed in a laying barn. Roosters are known to be aggressive around other males and towards hens when mating. So to introduce so many of them to overcrowded confined sheds can only be described as cruel as the birds have no way of escaping each other. When older males become less productive breeders they are slaughtered and younger males are introduced who are more sexually active. This is called spiking a flock. The breeding hens really don't get a break and are repeatedly mated. Parent birds are raised and kept until approximately 64 weeks and produce about 160 fertile eggs and are then slaughtered. The fertile eggs are taken from their mother and are then incubated in hatcheries. The male chicks are useless to the egg industry and do not produce eggs so they are destroyed via maceration or gassing and the females are either moved onto a rearing site to become conventional egg layers or are sold to backyard hen owners. So as you can see these hens are all coming from the same process. To support this industry is to support horrific animal cruelty. Another point to mention hens have been selectedly bred to produce more eggs than usual. Naturally a hen might lay 10 to 12 eggs a year but because of the various ways these breeds are manipulated they can lay anywhere from 250 to 320 eggs per year. This puts an enormous strain on the hen's reproductive system. Because of this the hens will need to replace the vital nutrients that are lost. So instead of taking the bird's eggs for yourself feeding their eggs back to them is best. Now let's just say for argument's sake you rescued a hen from a factory farm to avoid the hatchery and breeder sheds. You only ate the inside of the egg and fed the shells back to the hens in your backyard and gave them extra nutrients to replace what they lost from laying. You didn't eat eggs at all outside of your home or in any other circumstance so you didn't support any other aspect of the egg industry. You were vegan in every other way. You didn't use wool, leather, silk or down.  This is a gas chamber. Eggs.  Like the guy I debated. You weren't eating cows or supporting the cruel dairy industry.  Even then I would still say that the way you are viewing these animals is the problem. This is where it all begins. You are still viewing eggs as a product. You are still viewing hens as a resource. The only reason factory farms exist. The only reason slaughterhouses exist is because human beings see an animal and they see a product. Instead of seeing an individual, they see something to gain. The only reason people have chickens in their backyard is because they want the eggs that come out of them. And what happens when they stop laying for you? You stop caring. You send them to slaughter. Human beings need to stop viewing animals as their own little personal slaves and stop making bad excuses to exploit them. Yeah, they're in my backyard. They're happy. You supported an industry that massurates male chicks. I'm my chickens happy. If you truly cared about animals, you would be vegan. Thanks for watching. Peace.