%0 Journal Article
%T Engineering RsDddA as mitochondrial base editor with wide target compatibility and enhanced activity.
%A Cheng K
%A Li C
%A Jin J
%A Qian X
%A Guo J
%A Shen L
%A Dai Y
%A Zhang X
%A Li Z
%A Guan Y
%A Zhou F
%A Tang J
%A Zhang J
%A Shen B
%A Lou X
%J Mol Ther Nucleic Acids
%V 34
%N 0
%D 2023 Dec 12
%M 37744175
%F 10.183
%R 10.1016/j.omtn.2023.09.005
%X Double-stranded DNA-specific cytidine deaminase (DddA) base editors hold great promise for applications in bio-medical research, medicine, and biotechnology. Strict sequence preference on spacing region presents a challenge for DddA editors to reach their full potential. To overcome this sequence-context constraint, we analyzed a protein dataset and identified a novel DddAtox homolog from Ruminococcus sp. AF17-6 (RsDddA). We engineered RsDddA for mitochondrial base editing in a mammalian cell line and demonstrated RsDddA-derived cytosine base editors (RsDdCBE) offered a broadened NC sequence compatibility and exhibited robust editing efficiency. Moreover, our results suggest the average frequencies of mitochondrial genome-wide off-target editing arising from RsDdCBE are comparable to canonical DdCBE and its variants.