Journal
JOURNAL OF ZHEJIANG UNIVERSITY-SCIENCE B
Volume 22, Issue 1, Pages 73-86Publisher
ZHEJIANG UNIV
DOI: 10.1631/jzus.B2000282
Keywords
CRISPR/Cas9 genome editing; Double-strand break (DSB) repair pathway choice; Target binding affinity; Target residence
Categories
Funding
- National Natural Science Foundation of China [31671385, 31870806]
- Zhejiang Provincial Natural Science Foundation of China [LY18C050001, LQ20C050004]
- Fundamental Research Funds for the Central Universities in China [2019QNA7031]
Ask authors/readers for more resources
The CRISPR/Cas9 technology is widely used for genome editing, with the variation in target binding affinity and residence time potentially affecting the choice of DSB repair pathway and leading to heterogeneous mutations. This presents an opportunity for optimizing Cas9-based technologies.
The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is widely used for targeted genomic and epigenomic modifications and imaging in cells and organisms, and holds tremendous promise in clinical applications. The efficiency and accuracy of the technology are partly determined by the target binding affinity and residence time of Cas9-single-guide RNA (sgRNA) at a given site. However, little attention has been paid to the effect of target binding affinity and residence duration on the repair of Cas9-induced DNA double-strand breaks (DSBs). We propose that the choice of DSB repair pathway may be altered by variation in the binding affinity and residence duration of Cas9-sgRNA at the cleaved target, contributing to significantly heterogeneous mutations in CRISPR/Cas9 genome editing. Here, we discuss the effect of Cas9-sgRNA target binding and residence on the choice of DSB repair pathway in CRISPR/Cas9 genome editing, and the opportunity this presents to optimize Cas9-based technology.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available