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- Nat Biotechnol 2020-07;
New base editors change C to A in bacteria and C to G in mammalian cells
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The coding sequences in E. coli for expression of AID15 and the evolved TadA variant (ABE7.10; ref. 8 ) and in human for expression of Ung, with Cas9 variant linking regions (C-terminal and N-terminal), were synthesized (GenScript)...E. coli vectors expressing Ung and AID with mammalian codons, and human vectors expressing Ung and APOBEC with E. coli codons, were synthesized and assembled to produce pAPOBEC-nCas9-Ung and pUng-nCas9-AID, respectively, by GenScript | |
AbstractCurrent base editors (BEs) catalyze only base transitions (C to T and A to G) and cannot produce base transversions. Here we present BEs that cause C-to-A transversions in Escherichia coli and C-to-G transversions in mammalian cells. These glycosylase base editors (GBEs) consist of a Cas9 nickase, a cytidine deaminase and a uracil-DNA glycosylase (Ung). Ung excises the U base created by the deaminase, forming an apurinic/apyrimidinic (AP) site that initiates the DNA repair process. In E. coli, we used activation-induced cytidine deaminase (AID) to construct AID-nCas9-Ung and found that it converts C to A with an average editing specificity of 93.8% ± 4.8% and editing efficiency of 87.2% ± 6.9%. For use in mam... More KeywordsMost Popular Services
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