Reunion! : A Follow-up with Dr. Karolin Luger and the Hosts of Biocast

In this episode, I co-hosted Dr. Karolin Luger with Chinmay and Parth, the co-hosts of a partner podcast with From where does it STEM? called Biocast. Biocast is a biology podcast made to serve as a resource for anyone interested in biology: whether it be about recent breakthroughs in research, common topics, or even just a general interest, they hope to encourage people to explore the world of biology and help them delve deeper into the wonderful science of life itself.I hosted Karolin in a previous episode back in 2021, and this is the first time I’ve done a follow-up with a previous guest! If you forgot, she is a scientist at the University of Colorado, Boulder who solved the structure of the nucleosome using X-ray crystallography. More recently, she was awarded the 2023 World Laureates Association Prize in Life Sciences or Medicine.

Transcription

Transcribed by Gracie Dellinger (she/her)

J: JP Flores

K: Karolin Luger

C: Chinmay

P: Parth

(Sometimes, I could not tell whether Chinmay or Park was speaking, so I tried my best!!)

J: Like I was telling I was telling them before the call. I was like, Karolin is really nice. You’re gonna be starstruck because they’re nervous before this. And I was like, Karolin is the perfect first person for this.

K: Yeah, please don’t be nervous. There’s no reason whatsoever. And like like, they said. If I use any cuss words we can edit them out. So that’s good. Cause I don’t want to be like the president of Harvard who gets.

J: Oh yeah.

K: That was horrible. Yep.

J: Well, can you first start? Can you first tell us about the wild stories of you, you know, being a badass in the wilderness? I know you’re going on vacation in a couple of weeks, so can you walk us through that? I’m excited for you. What are you up to? What are you doing?

K: Well, I’m actually not going to the wilderness, unfortunately. I used to be a badass after my PhD; I actually was with my husband. We took a year off, and we canoed down the entire Yukon River in our little rubber canoe and fished and lived off the land. It was really fun. No like. In a couple of weeks, I’m going to Europe to Stockholm to give talks and advisory boards and see my parents. So you know nothing super exciting.

C: That’s still awesome.

K: Yeah, that’s still good. To get out, especially to see my parents.

J: Yeah. Were your parents scientists?

K: No, no, my dad was an electrical engineer. My mom’s a stay at home, mom. My brothers are all super smart geniuses in physics and electrical engineering. And so I was always like the third, the youngest, trying to keep up just navigating the world of biology for the first time. Really on your own.

C: What was that like? Just navigating the world of biology for the first time. Really on your own.

K: you know. I didn’t have any guidance. I just love watching plants grow, and I planted a lot of seeds. I wanted to be a botanist, and so I really didn’t know anything about it. I had no formal training. My teachers were not that great, so you know, I just kind of had to make it up as I went along. There wasn’t the Internet, very sad.

C: Fair enough. I think you mentioned earlier how science wasn’t really accessible to women. Like, what advice would you offer to young women, young scientists who aspire to follow a path similar to yours, coming from a background of like physics and math. And now doing biology.

K: Yeah, I mean, just don’t listen too much to other people who tell you can’t do it and try to put you into a specific mold and then, really early on, try to find people who believe in you, mentors. Usually, what helps is if you find other people who are like you, whether you are a woman or that really helps because they can relate to the more specific problems that you might encounter. But really, you know, I’ve had some fabulous male mentors as well, who just realized that there was an issue there. So I think just, you know, listen to the right kinds of people, not to the wrong ones.

C: How do you know which ones are the right kinds of people, though?

K: Yeah, that is a super good question. I don’t know, I think. Just go with your gut feeling, and you know you know in your core what you’re capable of, and if you have little doubts, and people come in and reinforce those doubts. That’s probably not the best person to listen to. But it’s more people who will recognize those doubts and say, Yeah, realize this is a limitation. This is what you might be able to do to overcome this. Those are the right kinds of people.

P: Yeah, that’s a great advice. Don’t listen to the haters.

K: Basically, when I was a second-year undergrad, actually, like you. We were, a couple of my friends and I, were kind of ranting against the situation, and how science is really kind of closed to women. In a way they were not. There were no role models. And this professor basically said, Oh, I’ve had many female students saying the same thing. And now they’re found their true destination. They’re happily married, and they’re raising families. And we’re like, really like. And we didn’t know enough to report him, because at that time you didn’t really do that. But that really was not a good piece of advice. So don’t listen to these people.

J: Yeah. So, on the topic of mentorship, before I hit record, I mentioned how I applied to the HHMI Gilliam fellowship. What the hell is an HHMI investigator? So I know the HHMI Gilliam is a very mentor-centric type of fellowship where you know the advisor and the pair gets awarded the advisor can go to Hhmi and get, you know, training on mentorship, but I know that you are an HHMI investigator, which I think is different.

K: So it’s very different. Yes, so I don’t know whether you all have seen the movie the aviator with Brad Pitt; it’s about Howard Hughes, Mr. Hughes, and he. He inherited a lot of money and made a lot of money, was quite the visionary, was also mentally very ill. And when he died he left without a will, and so, in order to avoid to pay taxes. It’s my understanding that they started this HHMI program, and for a while, it really didn’t do very much, but then somebody took over, and they turned it into a program where you can apply, and they fund, allegedly, people, not projects. And so they they look for people who they think has the potential to move science forward. And what is really unique about this program, they don’t stick us all in a silo and have us all work together. They leave us at our host institutions, which I think is amazing because that way, the ripple effect is much larger. And you know, we’re quite well funded, and we can help others move forward, and we can kinda pass it along and and and promote that idea of scientific excellence. Now it’s it’s really quite competitive. And every 5 to 7 years we have to be evaluated. We go there. We have to present what we did the last 5 to 7 years. What kind of papers did we publish? What kind of what did our Mentees do? So they now place quite a bit of emphasis on mentoring as well. What are our former lab members up to now? Are they still doing science, or did they all disappear, disgusted it from this whole enterprise. And you also have to put forward some ideas for what you want to do with the next in the next funding period, and it has to be kind of visonary may be the wrong word, but it has to be novel. Can’t just do the same old, same old. You have to have some fresh ideas. So you know, it’s a really fantastic program, because it really allows me to pass it forward to my colleagues and and have a wider effect in the scientific enterprise.

J: Yeah, it sounds like a very fancy Schmancy title. So speaking about like.

K: No, actually, no, it’s really tries very hard not to be elitist. And I think part of it is really that they are leaving us where we are, it would be super elitist, I think, if you all whisked us away and put us into some gold-plated building with, you know, and but that’s not what’s happening. So I really like that.

C: I think that’s really cool, though, like it kind of started off as like a tax fraud kind of thing.

K: Yeah, we might not want to put that in the podcast. It is accurate. I’m not that familiar with the exact history of this whole thing, but that’s kind of how it started out.

P: To all the listeners listening. Make sure to pay your taxes. So you get evaluated every 5 to 7 years, like, what have those? What has that timeline looked like for you?

K: So I’ve just been through my third renewal. And yeah. So I’ve been. I’ve been doing this for quite a while. I’ve been super fortunate. and you know, I’m just the front person like I live and die with the people in my lab and their dedication and their hard work. And you know, all the way to the undergrads, you know, they all make important contributions. They all work really hard. They know what’s at stake. And so, you know, I’ve been lucky picking the right people who really contributed a lot to this.

J: Yeah, definitely. So you’ve shown that you’re a world-class mentor doing world-class science. And I think we have to congratulate you on winning the 2023 World Laureates Association Prize right in life science through medicine. Can you talk a little bit about what this means for you personally and professionally like, how does it impact your work moving forward?

K: So this is kind of, they also call it the Chinese Nobel Prize, and it was awarded for the work I did as a post-doc, so like almost like back in 1997 and it was awarded to my co-mentor, as well as to a person who became who did work before my mentor was even involved in this. And so we’re all awarded this prize together for work we did as post-docs, kind of building on each other’s expertise. And it’s for the structure determination of the nucleosome, which is now in every textbook. And to me, you know it was like, I’m a little ambivalent about prizes, because it can be a little arbitrary, if you will, because it takes a lot of time to promote people to put them forward for prizes. It’s actually a lot of work on behalf of the nominators. And so, if you don’t have a strong believer in you, then you don’t really get nominated, even if you do really fabulous work. But having said that I was super happy for for the nucleosome, which I think is like you know, piece of of common knowledge and and part of the of the science landscape. And I was really happy that the nucleosome of got this prize, shall we say? And I? Really it was like the awardee of this, of this whole prize. It was really fantastic. I went there and I didn’t know what to expect. And they, I mean, they treat scientists like rock stars over there. I felt like Beyonce over there, so it’s really a lot of fun. And then, you know, after a week we had to come back and like, Oh, now I have to cook my own dinner again. That’s

J: Is that when you ask me for a clip from the podcast?

K: Yeah, yeah, that’s exactly what it is. Because I did kind of a general. They wanted background information that was relatable because they did a lot of press events there. And so it’s quite possible that a few quite a few people in China, listen to your podcast because, I really like that podcast. I think that was a lot of fun.

J: Me too. I listen to it a lot actually.

K: I’m not even sure what to say about that. I think you need to get a life, if that’s your if that’s your idea of entertainment. I’m a little concerned.

P: Oh, no, I was waiting for Parth. Yeah, you can go ahead. Go ahead. No, no, you got it. You got it. So I guess you talked a little bit about your journey like to the prize. But can you talk about your journey like as a scientist in general, like how you started off with your experiments? Like just the upward journey to finally winning the Nobel Prize of China.

K: Well, what can I say? I was trying to keep up with my brothers, and I really like I really like plants. And so I thought, well, you know, I could do botany, but that didn’t really sound like there were a lot of job opportunities. And so I went to the nearest university. And I studied microbiology, which was kind of the closest to this vague idea of molecular things I wanted to work on. And you know I was not… It wasn’t the greatest. They were really focused on taxonomy. And so we spent a lot of time counting warts on mushroom spores and things like that. But I did live. I did live right next to the biochemistry department. and at that time, biochemistry wasn’t even a major. And I started to go to their seminars, and you know it was horrible. I didn’t understand a single word I just said in the back, and I was like, Oh, my God! One of these days, I will understand all of this. I was just like riveted, and I really don’t can’t explain why, but they noticed me, and then they adopted me in their lab, and they and I started to to work as an undergrad in their lab doing, you know, just everything was exciting, like making a DNA Mini prep, and running a gel. And you know. Then, you know, I left the lid off a centrifuge and almost broke it, and it’s just like the whole thing is just very exciting. And that’s kind of how I got hooked. On doing research, nothing worked. It was really quite frustrating. And then I went to do my PhD; at that stage, I don’t think I contemplated much about whether I should do anything else. I just was kind of seeing the obvious choice and maybe people suggested to me that I could do it. And you know, that’s kind of this subversive mentoring that I got possibly that people thought like, Yeah, you might be quite good at it. I will say, I think, looking back now, I have a lot of undergrads in my lab, and there’s some that are very intense, and I really see myself in them. I think I was kind of a pain in the rear because I was, like, so focused on intense and, like, wanted to know everything. So then I did my PhD, and that work, really well, I’ve I had a really strange projects, a very creative project, just like, Look. do something and see what happens. I actually made the first circularly permuted amino acid sequence because at that time we didn’t really know when something gets translated by a ribosome whether it matters what part comes out. And so what I did is just fuse when you see an end termini and cut it open, at another place, and made a gene like that. and study the folding of this protein. And people are like, Why do you want to do this? This is never going to work. And it did work, and we could publish it, really well so that was really super fun and kind of one of these experiments raising like, why, really, why are you doing this? Just because you can, and you know, I think there’s a lot of merit to doing science like this. You come to all of your signs like this, but having the occasional pet project just to kind of poke it and see what happens.

P: Prove all of the haters wrong.

K: It’s not even that. It’s just like, you know. Where do the hypothesis come from if you don’t do weird experiments right? So then, like I said, my husband and I went on a year-long trip to cavort around the country and did not think about science much. Then I started a post-doc and I really wanted to learn X-ray crystallography. Which I found fascinating because we had this super boring lecture as a grad student, and I was just really fascinated by these little diffraction spots and how you get from there to a structure. So I thought, oh, just learn that. And I applied to this lab, and this professor said, oh, you know, you can just like solve the structure of the nucleosome. And we’re almost there. It’s almost like done. And that’s how you can learn crystallography, and then you can do something more interesting. And he basically didn’t tell me that this was one of the hardest projects in structural biology that existed at that time. So, I was like very blissfully ignorant. I said that sure, you know, like roll up my sleeve, let’s do it. Where do I start? So? So that took a long time. That took like 8 years. Really, really, really difficult project. And it was very different to the project I did as a grad student was very single-minded, you know. No creativity, just like throwing stuff at the wall to see what sticks. And eventually, you know, we managed to pull it off. And That then was a pretty big paper, and it gave me pretty good job prospects, I decided. We decided at that time that we’ve kind of had it with Europe. It was a little too close, and then I grew up there, so I should say, and it was a little too close-minded and then very male.Hang on. Check my audio device. Can you still hear me? Okay, good. So, it was pretty male-dominated at that time. And like I said, you know, women weren’t that common in faculty positions. So we went to the US. And actually made the of the deliberate decision rather than go to like one of the ivy leagues to go to a smaller undergrad institution, Colorado State University, where there was a very strong teaching mission and undergrads. And because I really like I didn’t like the Elitist types of institutions. We had to pay a lot of money to get an education so I worked there for 16 years, and I started my lab. With you know a lot of enthusiasm from on behalf of my colleagues. I had to start structural biology crystallography. At that time, I had to buy the generator and install all of that stuff. So that was really fun and then get my program off the off enrolling. I got queues. I got funding. And then, after, like, 16 years or so, it was getting a little stale, and you know, I still loved it up there, but I thought, like, let’s just try something. Mix it up a little. So, I moved like 60 miles down the road to Boulder, which is where I am now. And we’re still doing our thing. And but you know, ironically, when I arrived here, this is just when the resolution revolution in CryoEM took off. And so wouldn’t you know it? I had to start single-particle CryoEM here, and it’s kind of a Deja Vu all over again.

J: Can I quickly bring attention to one of your undergrads at CU Boulder? I believe one of them has a podcast. And I listened to it very recently. And in this podcast I learned that you really like the liver and liver function. I think he wrote a song and like played it for you in the class, or something like that.

K: Oh, yeah. Yeah. Oh, yeah, of course. Yeah. That was, so I used to teach metabolism and human disease. and it’s a pretty difficult class. And in the end, I used to give them the opportunity to get extra credit for creative activities. And that was before chat GPT. Now you can’t do it. It’s kind of pointless. But at that time, that particular class was very creative, and I had songs and skits and ballads, and these guys wrote an original song about the lonely liver. Oh, that that the selfless liver! Because I make this point in my class that the liver is kind of the Amazon fulfillment center of the body. And there’s a lot of things going on, and it just gives and gives and really doesn’t keep anything for itself. so they made a very sad and beautiful song about that. J: Yeah, it’s so good. If they’re still around, tell them that they should go on American idol if they want to come to UNC, give them my email for med school.

K: I think I know how to find them. And I agree. And I think, the woman who sang with him, I think she actually is a songwriter. In addition, I think, actually, she wrote that song. But I’m not 100% sure anymore.

J: Well, it’s clear that you have a strong passion for teaching undergraduates, and I know we’ve had this conversation before. But why do you love teaching so much? I think you mentioned that it was. It was the feeling of being able to help others. But has that evolved at all like, what is your answer now?

K: Well, I think it just comes from my own history. I’ve experienced what difference it can make how your teacher teaches you, whether it’s at the high school level or the undergrad level. I think science is fascinating, and more often than not it’s kind of being destroyed by how it is being taught. So, I just really like, I just love science. I love the process of discovery. And that’s really what I wanna pass along to students. Now, we’re kind of in this teaching revolution. I think it. It’s I don’t know whether you guys still have a lot of like normal traditional classes or whether you have mostly the flipped classroom and the active learning. I think there’s kind of a change in how we think about classroom teaching. That’s, for me, a little harder to keep up with. But I just think it’s it’s I think the difference you can make in a freshman or a second-year student by introducing them to the process of doing. Science can be life-changing. And I really, I think that’s good. You know, why not affect somebody’s life in a good way if you can?

J: Right?

C: Yeah. Speaking of being taught in the classroom and such, throughout our undergraduate molecular biology classes Parth and I are only second years, so we’ve only taken so many. But I’m sure JP has taken many more, and you as well. But it was emphasized, you know, that prokaryotes don’t really have histones, but you just published a paper in Nature actually titled Histones with an unconventional DNA binding mode in vitro are major chromatin constituents in bacterium Dellavibrio bacteriavores. First of all, first of all, that species name sounds like a Harry Potter spell like I have no idea.

K: You have no idea, this thing is so cool. But did you have a specific question about that?

Other: Well, can you just give us, you know us and our listeners, a little bit more insight on this and what it really means for biology moving forward as well?

K: Yeah. So, I try not to overstate it because we’ve really just only scratched the surface. But when you look at any molecular machine in our bodies, you want to know who invented it. Where did it come from? You usually go back to the primitive organisms. And for eukaryotes, that would be, for example, Giardia. You know the causative agent of beaver fever, some other single-cell organisms that are arguably kind of low on a totem pole. And if you look at the nucleosomes and the machinery that deals with them, it’s very complicated, and it’s all already 100% there in those organisms. And so you can’t really look at the evolutionary history seems like before we even decided to eukaryotes, we’ve had to have a pretty good system in place already to organize our genomes because eukaryotes have these massive genomes. You’re really bad at housekeeping. So it’s just you just need some organizing principles. So there’s this theory that the first eukaryote came into being by a bacterium eating an archaeon. So archaebacteria and the Archaeon gave rise to the nucleus, and the bacterium kind of gave rise to the rest. And then a lot of other unexplained magic happened. And, Tada, here we have our eukaryote, but we’ve worked on archaeological histones, and arguably they are the precursor of our own histones. Still, there are a lot of unexplained phenomena. And so we started to poke around in bacteria mainly it was a Covid project, and everybody was stuck in front of their computers. And so you could just do a lot of genome browsing, which is, my students did. And we started to look at all the giant viruses, and we found histones in all of these organisms. So, this particular organism, Dellavibrio bacteriavorus. I don’t know whether you guys speak Latin, but he literally means bacterium-eating bacterium. So, this guy, this guy is amazing, is very small, and he swims around and he finds himself a juicy fat E coli to eat. And it burrows itself into the outer cell wall, and most bacteria have 2 cell walls. They have an outer and inner one, and so that in between there’s a space. And so this guy lives in between. In that space, it closes the door behind him, which is fabulous, and it has some really interesting structural implications. It lives in there protected. Then it makes a lot of enzymes. It puts them into the cytoplasm of its prey, and the enzymes just digest the prey bacterium, the nutrients out. It sits there in this nice, protected space, and it uses all the nutrients to make new bacteria. And so this guy has histones, and we don’t know they bind DNA very, very differently, so they shift the whole paradigm of how we think about his phones. We don’t know where it got it from. It might have stolen it from some eukaryote and just used it for a different purpose, mutating it a little bit to change the binding mode for its own nefarious purposes. We are not really sure. But in a way this bacterium is almost like a virus. Speaking of viruses, we also work on giant viruses. And these things are literally, they’ve only been discovered in the 2000s. They have been detected so late because, traditionally, what you use to isolate viruses, you filter bacteria, or whatever lysates, and then whatever goes through is small. That’s a virus, and whatever is large is a bacterium. And so these things are so large that they kind of look like bacteria. And they have histones, and they’re honest-to-God viruses. They are also quite bizarre. They are large because their host organism is amoeba, and amoebas like to eat bacteria. So these viruses pretend to be bacteria, and so they just get eaten. Then they just get killed by whatever they are trying to eat. So it’s like a Trojan horse kind of thing.

J: That’s amazing. Is it like a histone optimer type? Or how many histone proteins do you see? Are they?

K: Yeah. So, for the Archaea and for the bacteria, we just see a single histone. And it’s a very minimalist histone. The giant viruses have all 4 histones, but some of them have them fused in either a single chain or in 2 different chains, so they fused them together. They’re also very badly conserved. They don’t. They’re not similar at all to the host histones. So it’s actually really quite fascinating. And also, incidentally, you might have seen my collaborator. They’re going around and digging up permafrost and you might have heard of the zombie virus. So they’re actually reviving giant viruses that are 40,000 years old, which is just amazing.

Other: Also, terrifying.

K: You know, it is, in a way. But they really do infect mostly aquatic life. But yeah, you’re not wrong it is very true.

Other: Yeah, just the fact that we can do that.

K: But these are the types of viruses that we work with. Unfortunately, the zombie viruses. Those particular ones don’t have histones, which really bums me out because I would have loved to work on them. But we’re constantly looking around for weird critters that might have histones. And how to look at how they use them?

Other: Yeah. So you’ve obviously done a lot of lot of interesting work, a lot of work in general. With the nucleosome. You mentioned the histones now. On your Wikipedia page, it’s highlighted that you solve the 3D structure of chromatin, which is kind of like a superhero of the cellular world. Yeah. So, could you talk a little bit about how you got involved with the molecular superhero and how you got the idea? For projects in the first place, the implications like you predict on that discovery, like having and what it’s already had, and most importantly like how it feels to have a Wikipedia page on yourself.

K: Well, I know everybody can make a Wikipedia page. It’s not. I think I think I don’t know. I don’t think that’s such an amazing accomplishment. But, actually, when I started to work on chromatin and nucleosomes, I thought they were really boring. And at that time, they were actually considered to be very boring. All the cool kids, we’re working on transcription factors and TDP and PF2 and all these kinds of things. That was kind of the big thing, activators. And everybody thought nucleosomes were very boring. Now, I just really wanted to learn structural biology, because I’m a very visual person. And I and to me, that’s just I just love to do science that way. And you know, everybody’s different, like cell biology. I tried that as an undergrad. I just thought it was very boring. I just really like to look at atoms. What can I say? And so and so then, for the longest time, I still didn’t think it was that interesting. While we were in the process of making that discovery or working on this project, the community independently realized how important these things were, so the timing was really good. In terms of making that big discovery, you know, it’s really cool. Like, you guys are also in labs and JP, also. So you are the expert. You’re the only people who know a certain thing. It’s true, though, like from your project, nobody else knows what you know. So that is kind of cool, right? And but, I didn’t, actually didn’t know that was the case. So, strangely enough, when I gave a seminar my first seminar about this big structure that nobody had seen, I flew to Chicago, and I was invited to give a talk there. I stayed up during the whole 10-hour flight because I thought, oh, my God! What if they asked me about this amino acid? What about if they ask me about that? It’s like they wouldn’t even know what to ask because I was just terrified. That hadn’t really occurred to me that they wouldn’t really know. Because they don’t didn’t have the information. So, you know. I really like the process of discovery, probably as much as the result. Just designing the experiment and trying to trick the system so it can observe the unobservable and you know the brain like one of you. I think Parth is interested in Neuro. You know that that is just a tremendously difficult problem. And I think we have to devise methods and tricks and approaches and think out of left field, how we can actually observe what’s happening. So, it’s really that part of the process that intrigues me more than the actual discovery. To be honest.

JP: Yeah, how do you train parts to do that?

K: I have 2 messages that I want everybody who’s engaging in science to know first. If you, if most of the time your experiments don’t work, or they give you unexpected results. Right? And it’s like, not what expected. And your gut reaction is like, that’s just horrible. Like I have to redo it. I have to do it over again. And my mantra is like, sit down and think about what this, what this could tell you, apart from the fact that you don’t know how to pipette, which you know sometimes that is, that is the case, or you forgot to add something. But let’s say you forgot to add something, and you see a weird observation. Don’t you say like, well, that sucks. I gotta do it again. But say, like, Well, okay, I get this really weird result because I forgot to add ATP or magnesium, or whatever. And look what I find, could that be something? And so, in a lot of cases. A lot of really seminal discoveries have come from this. Ha! That’s weird. I wonder what’s going on here? Rather than you know, feeling bad and just throwing it away and moving on. My students always laugh at me because I keep telling them that your experiment is talking to them. You just have to listen right to tell you something. It doesn’t just tell you you’re a failure. It tells you something important, you know, sometimes. Imagine, like, 99% of the time, it just tells you garbage. But like that, one time it tells you something important. You didn’t listen. That would be horrible, right? So that’s one of my mantras. It’s almost like making lemonade, right, from lemons. And then the second is, you know, a lot of my undergrads. They want to work on drug design, cancer, or Alzheimer’s. And you know that’s all good and laudable. But I do think you need to leave room for studying strange systems. And if you look back to, for example, the discovery of CRISPR, the CRISPR genome editing system, if you look back to restriction, enzymes, penicillin. A lot of these discoveries, I would say. Actually, the majority of transformative discoveries have come from people just poking around in weird systems just out of pure curiosity. Without like having the goal in mind of curing cancer. So leave a little room for that as well. And also, it’s like wicked fun, honestly. And also, added Bonus. It’s not quite as competitive as some of the other projects might be.

J: Yeah, I’m curious about finding funding for innovative ideas like that, you know. I think that’s the field of what? I’m blanking on it. Not natural. It’s natural product research, right? It’s

K: You know, it’s not even that. But you raise a good point about funding for these kinds of things. Nobody will give you money, right? So that is a problem. Yeah, so you know, you’d start with small pilot experiments that just poke around a little, see whether you can get a hook and then see whether you can get some data. And then you convince somebody that this actually might be interesting. Yeah. And you know, I agree it is very hard. And so I’m really fortunate to have funding from HHMI because they let me do this kind of stuff. They just want it to be interesting.

J: Or we could just join your lab and you could fund us.

K: Hey, you know, absolutely. Just apply, of course!

C: Would you say that the funding process, coming from someone who isn’t from your lab or doesn’t have the HHMI. Would they have to rely on getting lucky with the amount of small funding that they have, or small like stuff that they have, and then just moving from there?

K: Yeah, for sure. It’s luck. It’s also instinct, or it’s really hard to describe. And you know nobody has a perfect success rate, and I don’t either. We’ve had projects that were complete duds, of course. Right? When something really grips your imagination. And you really have that got feeling like, Oh, my God, that might be onto something! Then, you know, that’s a good time to read a little more to talk to a couple of people you might be. You might have to be careful not to give too much away, you know. Sometimes things can be a little competitive, and you know, more often than not, like, Oh, yeah. Well, that can’t work because it’s been done already, or somebody, you know you think it through. But then you can actually go. We started a lot of cool experiments and cool lines of research with undergrads just poking around a little because, for them, even if it fails, they still get good training. I’m not jeopardizing their PHD. They still learn the basics of science, and so that’s a relatively cost-effective way of doing business. It is. I’m not going to lie. It’s hard because funding agencies, especially the Federal ones, are very conservative, and they have to be because it’s taxpayer money, and they can’t be viewed as being frivolous. So that’s a bit of a conundrum.

C: And even with a lot of like the Federal funded agencies like compared to this, is, this is a bit more political, I guess, but like compared to some of the other branches of the government, or like other subsidiaries like, they don’t get as much money at all.

K: Yeah, we won’t mention any other funding any other agencies.

C: That’s military.

K: It is very true. And you know, admittedly, I think we’re the US is pretty good. And there’s actually more realization to fund that kind of basic research that I’m talking about as well. However, at many universities. For some reason, they go the opposite way, and everything has to be translational, which I think is a huge mistake. and I hope they kind of start to see the light, and everything has to be like small business and translational, and to the clinic. And I just I think that there’s a lot of need for that. But there’s also a lot of meat which is the pure curiosity-driven projects

C: awesome.

J: I’ve just been riffing. So Karolin, what do you think is next for the field of structural biology? So you know, we’re moving into. Yeah, nnon-eukaryotic stuff. Maybe we’re trying to play around a little bit. But what do you think is next, I know, like for me in the transcription field. We’re still trying to figure out the chromatin structure. And we need structural biologists for that. We don’t know how many answers were. But in in your little niche of structural biology. What do you think are the big, big things? Is it RNA Pol 2?

K: You know? I think that’s pretty much being done. There’s a number of fabulous labs that are making progress in leaps and bounds. It’s really quite impressive. I think generally looking at looking at these complexes. observing them at work, doing their job in their natural context. That is like dynamite. Imagine you could if you could just watch a polymerase doing its thing in the cell. That would be amazing, right? And I, truly, the field is moving in that direction. And so especially the field of cryo electron tomography, where you can actually thin slice cells and then put them under the microscope and see some quite astonishing resolution. So that’s really where things are is things are going. The other really important aspect that’s being lost by those static approaches like CryoEM and crystallography. That. They’re static pictures, and we really want to see things in movement. And now there’s approaches where they’re called single-molecule approaches, where you can like string them up between magnetic beads and then watch them do their thing. That’s a very artificial system. It provides a lot of really important information. But it’s a very artificial system. But that’s why I’m saying, if you can get the movement, the dynamic aspect into structural biology. That would be dynamite. And I think we’re gonna get there. I really do.

P: What do you think the timeline for something like that looks like.

K: you know, it’s so hard. I have no idea. There are a lot of really, really smart young people making amazing progress like, for example, I would. I would never have imagined that we can just plug in amino acid sequence and get really a really pretty good prediction of a fold out to me. That is amazing. And that’s really been transformative also. So you know, I have no idea what you guys are all capable of I think the sky is the limit, like, I’m not part of this anymore, because, I’m not the future. But you guys are the future right?

241 JP: Present? Right? Like, like, I think you are giving us a voice and platform, and you are inspiring us to be the future of science. So I do think that there is a place for you. And you are still doing wild stuff.

K: We are still doing wild stuff, and thank you for that because I really do aspire to inspire the next generation. But we’re not. We’re not pushing the technology forward. I think that’s really not something that we’re good at. We’re applying methodologies. And we are looking in unexpected places. Granted. And we’re finding unexpected things. But we’re not really pushing microscopy forward. For example, instrument development and computational development. You know, there are people who write software that allows us to do things with our raw data that haven’t been possible before. So that’s what I’m talking about. And I think it’s just like, this, this fearlessness that that you guys have your young people have like, oh, yeah, let’s just do it like, why not? That kind of goes away as people get older. And so that’s why I’m saying, like, you know, you guys are the future. Because you don’t know how you don’t really care that much if it doesn’t work right.

C: I think the future still needs current, excellent present mentors, and you’re absolutely one of them.

K: So, I’m not obsolete yet.

J: Yeah, I wish I went back to school, and if I could do it over again, I’d actually probably do biomedical engineering and data science. Because I think the skills you are learning in both of those are versatile enough to handle computers, and you’re versatile enough to go to medical school. You’re versatile enough to create new technology. If I could go back and I and do it again. I would actually.

K: But you can, like, you know, I’m really opposed to this, you know, boxing yourself in like you can absolutely do a post-doc in biomedical engineering. Why the heck not? The boundaries are not nearly as solid. And this is the other piece of advice that I would give. You guys, you know, don’t feel like, Oh, like, this is my major. And now I have to like walk on that path. And I think actually, the best science comes from people who really kinda look over the fence and say, like, Huh, let me see what’s over there. I can talk to people over there, and if you think back to the old days of Watson, Crick, Rosalind Franklin, and all these people. They’re all physicists and chemists, and they were embarking on this wild, squishy biology thing, and they came out of left field. So I think there’s really a lot of merit in that as well.

C: There’s some pretty crazy discoveries as well.

K: One or two important ones, I guess. I’ll give them that.