| Autor: |
Nguyen Tran, Minh Thuan, Mohd Khalid, Mohd Khairul Nizam, Wang, Qi, Walker, Jacqueline K. R., Lidgerwood, Grace E., Dilworth, Kimberley L., Lisowski, Leszek, Pébay, Alice, Hewitt, Alex W. |
| Předmět: |
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| Zdroj: |
Nature Communications; 9/25/2020, Vol. 11 Issue 1, pN.PAG-N.PAG, 1p |
| Abstrakt: |
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. Off-target effects and the feasibility for AAV-mediated delivery are the major barriers impeding the clinical in vivo application of base editors. Here, the authors report the small size AAV-deliverable Cas9-ABE variant that has improved on-target editing efficiency and reduced RNA-off target footprint. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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