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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, MP 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, MP (Mr Minh Thuan 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: | 16 |
| Deposited By: | Menzies Institute for Medical Research |
| Deposited On: | 2020-09-29 |
| Last Modified: | 2020-10-16 |
| Downloads: | 12 View Download Statistics |
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