// May 14, 2026 | American Society of Gene and Cell Therapy (ASGCT)
Next‑generation enzymes for targeted gene insertion
David Wiley LinkedIn
Director, Nucleic Acid Technology
Overview
This presentation showcases ElevateBio's comprehensive targeted large gene insertion technology, spanning novel large serine recombinases (LSRs) and R2 retrotransposons. From a diverse catalog of over 10 billion proteins, we identified 100+ active LSRs and active R2 retrotransposons. We achieved robust, specific insertion in primary human T cells, demonstrating a promising targeted insertion platform for in vivo or ex vivo therapeutic application. We are now leveraging generative AI to make large gene insertion fully programmable by designing purpose-built LSR proteins for specific genomic targets.
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My name is David Wiley. I work at Elevate Bio. We’ve developed a targeted large insertion technology that can meet a lot of the partners’ needs for insertion. Some of this could be in the CAR-T space, but we have specific enzymes that can meet a lot of the partners’ potential needs.
So we previously were working on CRISPR HDR based approaches for targeted large insertion using both viral templates and then non-viral templates. And we’ve added the large serine recombinase technology to our toolbox as this enables insertion of very large cargo without exposed double stranded breaks. So this makes it more amenable for allogeneic and more complex insertions, which is where the field is going to be going as well. So, to identify LSRs, we leverage our sequence catalog and pulled over 30,000 different candidates to capture the diversity of the LSR landscape.
We observed that over 80% of our LSRs are active, and many of these LSRs identified demonstrated higher activity than the literature benchmark. We have gone on and characterize these LSRs and looked at their integration profile, both where they’re integrating and how efficiently they’re integrating. We observed a huge range of diversity, from no integration all the way up to greater than 90% in some cell types.
And so we’ve characterized these in therapeutically relevant cell types and have identified leads that have favorable insertion profiles and robust efficiency. And we’re leveraging our CRISPR technology as well, so that we can enable a very modular platform to install landing pads precisely where we want. We’ve done this proof-of-concept work in primary T cells and have got installation of attB landing pad at greater than 90% at the TRAC locus. And then when we add our LSR components, we’re able to get over 67% installation of a CD19 CAR and across multiple donors. And this is under the endogenous TRAC promoter.
To date, we’ve identified over 100 active LSR and are continuing to expand this data set. And we’re applying our advanced protein engineering capabilities, including AI approaches, to further improve the specificity, potency and overall performance.
This technology is going to allow insertions of large cargo without double stranded breaks, will allow multiplexing in the ceiling for this technology, the efficacy potency ceiling is going to be much higher than with other technologies. So, we think this is going to be transformative for the targeted large insertion field.
About the Author
David Wiley
Director, Nucleic Acid Technology
David Wiley is the Director of Nucleic Acid Technology at ElevateBio. His work focuses on guide RNA engineering, mRNA engineering, and production optimization, and he is currently leading the development of next-generation large gene insertion technologies, focusing on R2 retrotransposons and large serine recombinases technology.
Before ElevateBio, David was at Prime Medicine, where he led efforts in pooled screening platform development and early mRNA optimization. Prior to that, he was at Vertex Pharmaceuticals, working in the Functional Genomics group to identify potential therapeutic targets using CRISPR-based pooled screening approaches.
David earned his B.S. in Cell Biology and Genetics from the University of Georgia and his Ph.D. in Cell and Molecular and Developmental Biology from the University of North Carolina at Chapel Hill. He completed his postdoctoral training at Boston Children’s Hospital.
In his free time, David enjoys carpentry and spending time outdoors.
