Chemical reprogramming enhances homology-directed genome editing in zebrafish embryos

Yagiz A. Aksoy, David T. Nguyen, Sharron Chow, Roger S. Chung, Gilles J. Guillemin, Nicholas J. Cole, Daniel Hesselson

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)
50 Downloads (Pure)

Abstract

Precise genome editing is limited by the inefficiency of homology-directed repair (HDR) compared to the non-homologous end-joining (NHEJ) of double strand breaks (DSBs). The CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 system generates precise, locus-specific DSBs that can serve as substrates for HDR. We developed an in vivo visual reporter assay to quantify HDR-mediated events at single-cell resolution in zebrafish and used this system to identify small-molecule modulators that shift the DNA repair equilibrium in favor of HDR. By further optimizing the reaction environment and repair template, we achieved dramatic enhancement of HDR-mediated repair efficiency in zebrafish. Accordingly, under optimized conditions, inhibition of NHEJ with NU7441 enhanced HDR-mediated repair up to 13.4-fold. Importantly, we demonstrate that the increase in somatic HDR events correlates directly with germline transmission, permitting the efficient recovery of large seamlessly integrated DNA fragments in zebrafish.
Original languageEnglish
Article number198
Pages (from-to)1-9
Number of pages9
Journal Communications Biology
Volume2
DOIs
Publication statusPublished - 23 May 2019

Bibliographical note

Copyright the Author(s) 2019. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

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