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Docking sites inside Cas9 for adenine base editing diversification and RNA off-target elimination

Nat Commun .. 2020-11; 
Shuo Li , Bo Yuan , Jixin Cao , Jingqi Chen , Jinlong Chen, Jiayi Qiu , Xing-Ming Zhao , Xiaolin Wang , Zilong Qiu, Tian-Lin Cheng
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Codon Optimization Human codon optimized deaminases TadA and TadA* were synthesized commercially (Genscript). Get A Quote

摘要

Base editing tools with diversified editing scopes and minimized RNA off-target activities are required for broad applications. Nevertheless, current Streptococcus pyogenes Cas9 (SpCas9)-based adenine base editors (ABEs) with minimized RNA off-target activities display constrained editing scopes with efficient editing activities at positions 4-8. Here, functional ABE variants with diversified editing scopes and reduced RNA off-target activities are identified using domain insertion profiling inside SpCas9 and with different combinations of TadA variants. Engineered ABE variants in this study display narrowed, expanded or shifted editing scopes with efficient editing activities across protospacer positions 2-16.... More

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