High-throughput functional variant screens via in vivo production of single-stranded DNA
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Show full item recordAbstract
<jats:title>Significance</jats:title>
<jats:p>We report a methodology for the pooled construction of mutants bearing precise genomic sequence variations and multiplex phenotypic characterization of these mutants using next-generation sequencing (NGS). Unlike existing techniques depending on CRISPR-Cas–directed genomic breaks for genome editing, this strategy instead uses single-stranded DNA produced by a retron element for recombineering. This enables libraries of millions of elements to be constructed and offers relaxed design constraints which permit natural DNA or random variation to be used as inputs.</jats:p>
Date issued
2021Department
Massachusetts Institute of Technology. Research Laboratory of ElectronicsJournal
Proceedings of the National Academy of Sciences of the United States of America
Publisher
Proceedings of the National Academy of Sciences
Citation
Schubert, Max G, Goodman, Daniel B, Wannier, Timothy M, Kaur, Divjot, Farzadfard, Fahim et al. 2021. "High-throughput functional variant screens via in vivo production of single-stranded DNA." Proceedings of the National Academy of Sciences of the United States of America, 118 (18).
Version: Final published version