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Engineering RsDddA as mitochondrial base editor with wide target compatibility and enhanced activity.


ABSTRACT: 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.

SUBMITTER: Cheng K 

PROVIDER: S-EPMC10514076 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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Engineering RsDddA as mitochondrial base editor with wide target compatibility and enhanced activity.

Cheng Kai K   Li Cao C   Jin Jiachuan J   Qian Xuezhen X   Guo Jiayin J   Shen Limini L   Dai YiChen Y   Zhang Xue X   Li Zhanwei Z   Guan Yichun Y   Zhou Fei F   Tang Jin J   Zhang Jun J   Shen Bin B   Lou Xin X  

Molecular therapy. Nucleic acids 20230909


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 DddA<sub>tox</sub> homolog from <i>Ruminococcus</i> sp. <i>AF17-6</i> (RsDddA). We engineered RsDddA for mitochondrial base editi  ...[more]

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