Quantification of evolved DNA-editing enzymes at scale with DEQSeq

Research output: Contribution to journalResearch articleContributedpeer-review

Abstract

We introduce DEQSeq, a nanopore sequencing approach that rationalizes the selection of favorable genome editing enzymes from directed molecular evolution experiments. With the ability to capture full-length sequences, editing efficiencies, and specificities from thousands of evolved enzymes simultaneously, DEQSeq streamlines the process of identifying the most valuable variants for further study and application. We apply DEQSeq to evolved libraries of Cas12f-ABEs and designer-recombinases, identifying variants with improved properties for future applications. Our results demonstrate that DEQSeq is a powerful tool for accelerating enzyme discovery and advancing genome editing research.

Details

Original languageEnglish
Article number254
Number of pages16
JournalGenome Biology
Volume24
Issue number1
Publication statusPublished - 6 Nov 2023
Peer-reviewedYes

External IDs

PubMed 37932818
ORCID /0000-0001-9335-9749/work/147143640

Keywords

Keywords

  • Base editing, CRISPR, Cas12f, Cre recombinase, Deep sequencing, Directed evolution, Site-specific recombination, DNA, Recombinases/genetics, CRISPR-Cas Systems, Directed Molecular Evolution/methods, Gene Editing/methods