Programmable C:G to G:C genome editing with CRISPR-Cas9-directed base excision repair proteins

Page view(s)
3
Checked on Jun 02, 2022
Programmable C:G to G:C genome editing with CRISPR-Cas9-directed base excision repair proteins
Title:
Programmable C:G to G:C genome editing with CRISPR-Cas9-directed base excision repair proteins
Other Titles:
Nature Communications
Publication Date:
02 March 2021
Citation:
Chen, L., Park, J. E., Paa, P., Rajakumar, P. D., Prekop, H.-T., Chew, Y. T., … Chew, W. L. (2021). Programmable C:G to G:C genome editing with CRISPR-Cas9-directed base excision repair proteins. Nature Communications, 12(1). doi:10.1038/s41467-021-21559-9
Abstract:
AbstractMany genetic diseases are caused by single-nucleotide polymorphisms. Base editors can correct these mutations at single-nucleotide resolution, but until recently, only allowed for transition edits, addressing four out of twelve possible DNA base substitutions. Here, we develop a class of C:G to G:C Base Editors to create single-base genomic transversions in human cells. Our C:G to G:C Base Editors consist of a nickase-Cas9 fused to a cytidine deaminase and base excision repair proteins. Characterization of >30 base editor candidates reveal that they predominantly perform C:G to G:C editing (up to 90% purity), with rAPOBEC-nCas9-rXRCC1 being the most efficient (mean 15.4% and up to 37% without selection). C:G to G:C Base Editors target cytidine in WCW, ACC or GCT sequence contexts and within a precise three-nucleotide window of the target protospacer. We further target genes linked to dyslipidemia, hypertrophic cardiomyopathy, and deafness, showing the therapeutic potential of these base editors in interrogating and correcting human genetic diseases.
License type:
Attribution 4.0 International (CC BY 4.0)
Funding Info:
This work is supported by Agency for Science, Technology and Research (A*STAR) and Industrial Alignment Fund Pre-Positioning (IAF-PP) grant H17/01/a0/012. Work by P.D.R. and W.L.C. is further supported by Advanced Manufacturing and Engineering (AME) Programmatic grant A18A9b0060.
Description:
ISSN:
2041-1723