Characterization of the interplay between DNA repair and CRISPR/Cas9-induced DNA lesions at an endogenous locus

Anne Bothmer, Tanushree Phadke, Luis A. Barrera, Carrie M Margulies, Christina S. Lee, Frank Buquicchio, Sean Moss, Hayat S. Abdulkerim, William Selleck, Hariharan Jayaram, Vic E. Myer and Cecilia Cotta-Ramusino ()
Additional contact information
Anne Bothmer: Editas Medicine
Tanushree Phadke: Editas Medicine
Luis A. Barrera: Editas Medicine
Carrie M Margulies: Editas Medicine
Christina S. Lee: Editas Medicine
Frank Buquicchio: Editas Medicine
Sean Moss: Editas Medicine
Hayat S. Abdulkerim: Editas Medicine
William Selleck: Editas Medicine
Hariharan Jayaram: Editas Medicine
Vic E. Myer: Editas Medicine
Cecilia Cotta-Ramusino: Editas Medicine

Nature Communications, 2017, vol. 8, issue 1, 1-12

Abstract: Abstract The CRISPR–Cas9 system provides a versatile toolkit for genome engineering that can introduce various DNA lesions at specific genomic locations. However, a better understanding of the nature of these lesions and the repair pathways engaged is critical to realizing the full potential of this technology. Here we characterize the different lesions arising from each Cas9 variant and the resulting repair pathway engagement. We demonstrate that the presence and polarity of the overhang structure is a critical determinant of double-strand break repair pathway choice. Similarly, single nicks deriving from different Cas9 variants differentially activate repair: D10A but not N863A-induced nicks are repaired by homologous recombination. Finally, we demonstrate that homologous recombination is required for repairing lesions using double-stranded, but not single-stranded DNA as a template. This detailed characterization of repair pathway choice in response to CRISPR–Cas9 enables a more deterministic approach for designing research and therapeutic genome engineering strategies.

Date: 2017
References: Add references at CitEc
Citations: View citations in EconPapers (4)

Downloads: (external link)
https://www.nature.com/articles/ncomms13905 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:8:y:2017:i:1:d:10.1038_ncomms13905

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

DOI: 10.1038/ncomms13905

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 ().