Timed inhibition of CDC7 increases CRISPR-Cas9 mediated templated repair

Beeke Wienert, David N. Nguyen, Alexis Guenther, Sharon J. Feng, Melissa N. Locke, Stacia K. Wyman, Jiyung Shin, Katelynn R. Kazane, Georgia L. Gregory, Matthew A. M. Carter, Francis Wright, Bruce R. Conklin, Alex Marson, Chris D. Richardson () and Jacob E. Corn ()
Additional contact information
Beeke Wienert: University of California
David N. Nguyen: University of California
Alexis Guenther: University of California
Sharon J. Feng: University of California
Melissa N. Locke: University of California
Stacia K. Wyman: University of California
Jiyung Shin: ETH Zürich
Katelynn R. Kazane: University of California
Georgia L. Gregory: Gladstone Institutes
Matthew A. M. Carter: Gladstone Institutes
Francis Wright: University of California
Bruce R. Conklin: Gladstone Institutes
Alex Marson: University of California
Chris D. Richardson: University of California
Jacob E. Corn: University of California

Nature Communications, 2020, vol. 11, issue 1, 1-15

Abstract: Abstract Repair of double strand DNA breaks (DSBs) can result in gene disruption or gene modification via homology directed repair (HDR) from donor DNA. Altering cellular responses to DSBs may rebalance editing outcomes towards HDR and away from other repair outcomes. Here, we utilize a pooled CRISPR screen to define host cell involvement in HDR between a Cas9 DSB and a plasmid double stranded donor DNA (dsDonor). We find that the Fanconi Anemia (FA) pathway is required for dsDonor HDR and that other genes act to repress HDR. Small molecule inhibition of one of these repressors, CDC7, by XL413 and other inhibitors increases the efficiency of HDR by up to 3.5 fold in many contexts, including primary T cells. XL413 stimulates HDR during a reversible slowing of S-phase that is unexplored for Cas9-induced HDR. We anticipate that XL413 and other such rationally developed inhibitors will be useful tools for gene modification.

Date: 2020
References: Add references at CitEc
Citations: View citations in EconPapers (5)

Downloads: (external link)
https://www.nature.com/articles/s41467-020-15845-1 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-15845-1

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-020-15845-1

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().