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Quantification of evolved DNA-editing enzymes at scale with DEQSeq
Lukas Theo Schmitt1,2, Aksana Schneider1,2, Jonas Posorski1
1Medical Faculty and University Hospital Carl Gustav Carus, UCC Section Medical Systems Biology, Dresden, TU Dresden, 01307, Germany.
Genome Biology
|November 7, 2023
Summary
DEQSeq, a nanopore sequencing method, accelerates genome editing enzyme discovery by analyzing full-length sequences, editing efficiencies, and specificities from thousands of variants simultaneously. This powerful tool aids in identifying superior enzymes for research and applications.
Area of Science:
- Genomics
- Molecular Biology
- Biotechnology
Background:
- Directed molecular evolution is crucial for discovering novel enzymes with enhanced genome editing capabilities.
- Identifying optimal enzyme variants from large libraries requires efficient and high-throughput screening methods.
Purpose of the Study:
- To introduce DEQSeq, a nanopore sequencing approach for rational selection of genome editing enzymes.
- To streamline the identification of high-performing enzyme variants from directed evolution experiments.
Main Methods:
- DEQSeq utilizes nanopore sequencing to capture full-length enzyme sequences, editing efficiencies, and specificities.
- The method enables simultaneous analysis of thousands of evolved enzyme variants.
- Application to Cas12f-ABEs and designer-recombinase libraries.
Main Results:
- DEQSeq successfully identified variants with improved properties from evolved enzyme libraries.
- The approach demonstrated efficiency in analyzing large-scale enzyme evolution experiments.
- Successful application to Cas12f-ABEs and designer-recombinases.
Conclusions:
- DEQSeq is a powerful tool for accelerating enzyme discovery in genome editing.
- The method significantly streamlines the selection of valuable enzyme variants for research and applications.
- DEQSeq advances the field of genome editing by enabling rapid identification of superior enzymes.
