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Engineering domain-inlaid SaCas9 adenine base editors with reduced RNA off-targets and increased on-target DNA editing

Citation

Nguyen Tran, MT and Mohd Khalid, MKN and Wang, Qi and Walker, JKR and Lidgerwood, GE and Dilworth, KL and Lisowski, L and Pebay, A and Hewitt, AW, Engineering domain-inlaid SaCas9 adenine base editors with reduced RNA off-targets and increased on-target DNA editing, Nature Communications, 11, (1) Article 4871. ISSN 2041-1723 (2020) [Refereed Article]


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Copyright Statement

The Author(s) 2020. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) http://creativecommons.org/licenses/by/4.0/

DOI: doi:10.1038/s41467-020-18715-y

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 Details

Item Type:Refereed Article
Research Division:Biological Sciences
Research Group:Genetics
Research Field:Genome structure and regulation
Objective Division:Health
Objective Group:Clinical health
Objective Field:Clinical health not elsewhere classified
UTAS Author:Nguyen Tran, MT (Mr Minh Nguyen Tran)
UTAS Author:Mohd Khalid, MKN (Mr Mohd Mohd Khalid)
UTAS Author:Wang, Qi (Miss Qi Wang)
UTAS Author:Walker, JKR (Miss Jacqueline Walker)
UTAS Author:Hewitt, AW (Professor Alex Hewitt)
ID Code:141152
Year Published:2020
Web of Science® Times Cited:1
Deposited By:Menzies Institute for Medical Research
Deposited On:2020-09-29
Last Modified:2020-10-16
Downloads:2 View Download Statistics

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