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Abstract

Precision genome engineering has dramatically advanced with the development of CRISPR/Cas base editing systems that include cytosine base editors and adenine base editors (ABEs). Herein, we compare the editing profile of circularly permuted and domain-inlaid Cas9 base editors, and find that on-target editing is largely maintained following their intradomain insertion, but that structural permutation of the ABE can affect differing RNA off-target events. With this insight, structure-guided design was used to engineer an SaCas9 ABE variant (microABE I744) that has dramatically improved on-target editing efficiency and a reduced RNA-off target footprint compared to current N-terminal linked SaCas9 ABE variants. This represents one of the smallest AAV-deliverable Cas9-ABEs available, which has been optimized for robust on-target activity and RNA-fidelity based upon its stereochemistry.

Item Type:	Article
Authors/Creators:	Nguyen Tran, MT and Mohd Khalid, MKN and Wang, Q and Walker, JKR and Lidgerwood, GE and Dilworth, KL and Lisowski, L and Pebay, A and Hewitt, AW
Journal or Publication Title:	Nature Communications
Publisher:	Nature Publishing Group
ISSN:	2041-1723
DOI / ID Number:	10.1038/s41467-020-18715-y
Copyright Information:	© The Author(s) 2020. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) http://creativecommons.org/licenses/by/4.0/
Related URLs:	Google Scholar: Hewitt, AW UTAS Profile: Hewitt, AW
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