Decorating chromatin for enhanced genome editing using CRISPR-Cas9

Chen, Evelyn; Lin-Shiao, Enrique; Trinidad, Marena; Doost, Mohammad Saffari; Colognori, David; Doudna, Jennifer A.

doi:10.1073/pnas.2204259119

Decorating chromatin for enhanced genome editing using CRISPR-Cas9

Journal Article · Thu Dec 01 23:00:00 EST 2022 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.2204259119· OSTI ID:2470930

^[1]; ^[1]; Trinidad, Marena ^[1]; ^[1]; Colognori, David ^[1]; ^[2]

University of California, Berkeley, CA (United States)
University of California, Berkeley, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

CRISPR-associated (Cas) enzymes have revolutionized biology by enabling RNA-guided genome editing. Homology-directed repair (HDR) in the presence of donor templates is currently the most versatile method to introduce precise edits following CRISPR-Cas-induced double-stranded DNA cuts, but HDR efficiency is generally low relative to end-joining pathways that lead to insertions and deletions (indels). We tested the hypothesis that HDR could be increased using a Cas9 construct fused to PRDM9, a chromatin remodeling factor that deposits histone methylations H3K36me3 and H3K4me3 to mediate homologous recombination in human cells. Our results show that the fusion protein contacts chromatin specifically at the Cas9 cut site in the genome to increase the observed HDR efficiency by threefold and HDR:indel ratio by fivefold compared with that induced by unmodified Cas9. HDR enhancement occurred in multiple cell lines with no increase in off-target genome editing. These findings underscore the importance of chromatin features for the balance between DNA repair mechanisms during CRISPR-Cas genome editing and provide a strategy to increase HDR efficiency.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH); National Institute of General Medical Sciences

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 2470930

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 49 Vol. 119; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Biological Sciences
CRISPR
Chromatin
Epigenetics
Genetics
Genome editing
Science & Technology

Decorating chromatin for enhanced genome editing using CRISPR-Cas9

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