NEB Podcast #64 -
Transcript
Interviewers: Lydia Morrison, Marketing Communications Manager & Podcast Host, New England Biolabs, Inc.
Interviewee: Nathan Tanner, Ph.D., Associate Director of Research, New England Biolabs, Inc.
Lydia Morrison:
Welcome to the Lessons from Lab & Life podcast, brought to you by New England Biolabs. I'm your host, Lydia Morrison, and I hope this podcast offers you some new perspective. Today, I'm joined by New England Biolabs' own Nathan Tanner. Nathan's here to talk about NEB's Enzymes for Innovation program, and how some of these enzymes are being used in new methodologies and applications. Nathan, thanks so much for being in the studio with me today.
Nathan Tanner:
Great, thanks for having me.
Lydia Morrison:
Yeah, I'm super excited to talk about the Enzymes for Innovation initiative with you today. So, can you give us an overview of what the Enzymes for Innovation program is, and what exactly are enzymes for innovation, and how they can be different from one another?
Nathan Tanner:
Sure. So, the Enzymes for Innovation, or EFIs, as we like to call them, really stem from us being scientists at NEB. We work in the lab with new enzymes every day, and our CSO, Rich Roberts, really had an interest in making more official the collaboration between our research labs and the outside world; to get these cool tools we find and make into the hands of people who can build technologies with them. So, as we study them in the lab, we find ones we think are really interesting and commercialize them, put them out as NEB products. Our normal products are geared around a specific application, something like doing PCR faster, a new ladder, new library prep, that kind of thing. But the EFIs are really unique. They do an activity that no other enzyme on the market does. We're not really sure what people are going to do with them, but we want to enable researchers and developers out there to get these enzymes, build new things with them, and just see what happens.
Lydia Morrison:
Why is it important to us that the scientific community be aware of the Enzymes for Innovation program?
Nathan Tanner:
Yeah, like I said, these are unique activities. No one has commercialized an enzyme that does the things that EFIs do. And in some cases, we can imagine a use for them; in some cases, we really aren't sure; but they're unique. And we're talented, great scientists at NEB, we like to think, but we're only a small piece of the biology and biotech world, but there are people out there with crazy ideas that really can't do the things they want to do because the enzyme that would help them doesn't exist. But if they check the EFI page, they can see all these cool new activities they're putting out and turn it into new technology.
Lydia Morrison:
So, how does NEB identify and develop these unique enzymes with functionality that we don't fully understand?
Nathan Tanner:
So, they all come from a research lab where the lab is the expert in that kind of activity or that kind of enzyme, and so they find one that does something well, does something interesting. And importantly, they're NEB products, so they're made with the same quality and expectations things people have for our products. But the enzymes need to be able to be expressed, be purified, be produced as stable, good NEB products, just like everything else; they just happen to have a selection of really weird and interesting activities.
Lydia Morrison:
Can you share some specific examples of Enzymes for Innovation enzymes that NEB has developed?
Nathan Tanner:
Yeah, sure. So, we have a whole collection across the whole portfolio of NEB enzymes of technologies, but a couple good ones. One of the first ones was a methylase, EcoGII. This is a non-specific n6A methylase, and people found good use for it in mapping chromatin, because the A's that are accessible outside of the chromatin can get methylated. The ones that are bound up in chromatin are protected. So, you can do mapping experiments with this enzyme, and that's proven to be really interesting for that field.
We commercialized a prokaryotic Argonaute from Thermus thermophilus. So, Argonautes are cool enzymes that are guided nucleases, like Cas proteins, but they're guided by DNA, not RNA. This one works at really high temperatures and it can cut specific sequences. So, it's a targeted nucleus, but it uses DNA not RNA, and works in a different way than the Cas9 that people are used to.
Another good one is an RNA ligase called RtcB. So, RNA ligases kind of lag behind a bit in the development compared to DNA ligases, but we're working on them every day, and this one has a unique ability to ligate ends that are different from the standard RNA ligase. It can handle a 3-prime phosphate or a cyclic phosphate, and these things are normally a deal-breaker for standard RNA ligases, but this one can handle those. So, there's a couple examples there, we have a bunch more, but these are the kinds of activities we try to put out to let people play with.
Lydia Morrison:
So Nathan, what are some of the most exciting or surprising applications of these enzymes?
Nathan Tanner:
It's great to see our customers buy these enzymes with cool activities and use them to build new technologies. A couple good examples are a repair helicase that we sell, Tte UvrD, and we don't have a lot of helicases in the catalog, but this one is unique; it can recognize any double-stranded DNA structure and initiate without a replication fork. Which is useful as a helicase, people can play with it, but it's found a lot of utility in suppressing nonspecific amplification, and diagnostic applications like LAMP and other isothermal amplification reactions.
We sell a flap endonuclease, FEN1. It's a structure-specific endonuclease that recognizes a 5-prime flap. So, whenever DNA has that feature, this nucleus can cut that out, uniquely targeted to that structure. And specific for industry workflows, we sell an immobilized T4 DNA ligase. It's everybody's favorite ligase, we've had it around forever, but this version is on magnetic beads. So, you literally pull the enzyme out of solution, you don't have to heat and activate it, it's really nice for automation workflows, and it's in the EFI bucket because enzymes on beads is new and weird, and we're not sure what people want to do with that, but it has really enabled some unique workflows with the ability to pull the ligase out of solution based on the beads.
Lydia Morrison:
Yeah, I'm curious if there are examples of ways that these enzymes have impacted research or industry practices.
Nathan Tanner:
Yeah, the diagnostic enhancements are really good ones. But I think that the real key is just enabling things that weren't possible before, so that people want to use a certain DNA, use a certain RNA, and they can't, because there's no enzyme that allows them to do it, or the natural state of that molecule is inaccessible. And we're really able to change that paradigm by putting out an enzyme that fixes that problem.
Lydia Morrison:
So, I know that NEB offers GMP-grade TelN. What is TelN, and why is that significant in the context of talking about Enzymes for Innovation?
Nathan Tanner:
Yeah, TelN is for our first what we call graduate. It started as an EFI and now has been promoted up the chain to a full NEB product, and now offered it from our GMP-grade class of products. And it's an interesting enzyme, it's what's called a protelomerase, but it cuts double-stranded DNA. It has a recognition site. It's a really long one, it's 50-something bases long, so it's a little bit different than a restriction enzyme, but it'll find its site and cut it.
What it does after that, though, is really weird. Instead of producing blunt double-stranded ends, it attaches the ends to each other. So, it makes almost... Think of it like a circle, where the ends are covalently attached to each other, which people had identified this in literature, we didn't discover this enzyme, but that was a really weird thing, and we were curious what people could do with it. It turns out it's really useful for making protected DNA molecules, and also when you make a lot of DNA, you can process it with TelN into individual pieces, and those pieces are then really useful as molecules to put into therapeutic applications, because they're protected, nothing can act on the ends. A lot of natural repair enzymes use the ends, and if you don't have ends, they can't do that.
So, TelN has found a lot of utility for producing therapeutic DNAs, and that's why we have promoted it up to our GMP manufacturing, because that's the quality of product that people need for those applications. So, it's a little sad to see it not be an EFI anymore. We'll always consider it our first graduate, but it's technically removed from the program just due to its success.
Lydia Morrison:
Yeah. You've used the word "graduate" a couple times. Can you explain the process and importance of graduating TelN?
Nathan Tanner:
Yeah, sure. So, we just made that up for what happens when an enzyme moves beyond EFI. Because, like I said, it's important that the EFIs are manufactured with the same quality performance expectations as all the NEB products, we just tend to make them at smaller scale, because we're not sure how many people are going to need it. We don't want to make so much that we could never sell it all or never get it out there. So, what we'll do is make a smaller scale, and then as the EFIs become more popular, people ask for them at higher concentrations, or in different formats, or just larger amounts, we see the need to make them at full product scale. So, that'll start to happen, and we'll look to put them in the catalog with multiple formats, maybe different quality expectations, like the GMP-grade.
So, really it just means it's beyond an EFI because we know what it's used for now. Somebody out there, or many people out there, have identified an application, are using that enzyme for that thing, and we know what the enzymes do and why we need to make a lot of it. So, graduation just means it's gone beyond EFI. We just make a lot more of it for a lot more people to use.
Lydia Morrison:
Yeah, and we also make it in GMP quality now, which is good manufacturing practices. How will offering the GMP-grade TelN benefit researchers in the biotechnology industry?
Nathan Tanner:
The GMP-grade products really are suited for therapeutic and regulated markets where there's a lot of traceability and documentation requirements on the production of those materials. So, NEB is not making an actual active pharmaceutical ingredient, but some of our customers can buy these enzymes and make DNAs or RNAs or whatever with those enzymes, and trust that all the documentation and quality is in place as they look to make these really highly scrutinized regulated products.
Lydia Morrison:
Yeah, so, super important, actually, in the use and development of the product in therapeutics. So, what future developments can we expect from the Enzymes for Innovation program?
Nathan Tanner:
Yeah, we're putting out new enzymes all the time, so it's always changing, there's always something new. So, we have a whole page on the NEB website where we list all the EFIs and any new ones. So, we'll continue to identify new activities. We have some coming out soon really focused around RNA modifications. There's a couple enzymes that do unique things to RNA. Lots of things coming out. Our labs keep identifying new enzymes, new activities, so stay tuned to see what we put out next.
But we aim to launch a few every year. The next one coming out is an enzyme called FTO, which is, again, an RNA modification enzyme. So, that'll be a fun one, we've been working on that for a while. And we're always looking for new ideas. We have an email address, enzymesforinnovation@neb.com, we'd love to hear what people want. If there's a format for one of the EFIs you're using that you'd like to see, let us know. And anything you're looking for. We can't promise to make everything somebody suggests, but a lot of times people are looking for something we already have, and if we get a lot of those requests, it'll incentivize us to put it in the catalog.
Lydia Morrison:
It's really cool to see how the enzymes that have come out of the Enzymes for Innovation program have been used to help develop other technologies, and I can't wait to see how they're used in the future. There are lots more Enzymes for Innovation out there for our listeners to explore themselves and to start playing with in their labs. And we'll definitely have links in the notes for this show to some of the resources that Nathan mentioned, and we certainly welcome feedback. So, thanks so much for being with us today, Nathan, I really appreciate it.
Nathan Tanner:
Yeah, of course. Always happy to talk about these cool enzymes.
Lydia Morrison:
Thank you for joining us for this episode of the Lessons from Lab & Life podcast. Please check out our show's transcript for helpful links from today's conversation. And as always, we invite you to join us for the next episode, when I'm joined by three Passion In Science Awards winners in the category of Arts and Creativity. These three individuals have used their knowledge of science to create some pretty incredible and unique forms of art. It's a don't miss episode, so please join us next time.
