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Engineered domain-inlaid Nme2Cas9 adenine base editors with increased on-target DNA editing and targeting scope

BMC Biol . . 2023-09; 
Ding Zhao , Xun Gao , Jiale Zhou , Jinze Li , Yuqiang Qian , Di Wang , Wenchao Niu , Tao Zhang , Mingyang Hu , Haoyang Xiong , Liangxue Lai , Zhanjun Li
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Gene Synthesis TadA-8e DNA fragments were then synthesized and cloned and inserted into diferent positions of Nme2ABE8e by GenScript Biotech (Nanjing, China). Get A Quote

Abstract

Background: Nme2ABE8e has been constructed and characterized as a compact, accurate adenine base editor with a less restrictive dinucleotide protospacer-adjacent motif (PAM: N4CC) but low editing efficiency at challenging loci in human cells. Here, we engineered a subset of domain-inlaid Nme2Cas9 base editors to bring the deaminase domain closer to the nontarget strand to improve editing efficiency. Results: Our results demonstrated that Nme2ABE8e-797 with adenine deaminase inserted between amino acids 797 and 798 has a significantly increased editing efficiency with a wide editing window ranging from 4 to 18 bases in mammalian cells, especially at the sites that were difficult to edit by Nme2ABE8e. In addit... More

Keywords

Adenine base editors; Inlaid domain; Nme2ABE8e-797; Nme2ABE8e-797Smu; Nme2ABE8e-797−C.