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

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-co...

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Detalles Bibliográficos
Autores principales: Cheng, Kai, Li, Cao, Jin, Jiachuan, Qian, Xuezhen, Guo, Jiayin, Shen, Limini, Dai, YiChen, Zhang, Xue, Li, Zhanwei, Guan, Yichun, Zhou, Fei, Tang, Jin, Zhang, Jun, Shen, Bin, Lou, Xin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2023
Materias:
Original Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10514076/
https://www.ncbi.nlm.nih.gov/pubmed/37744175
http://dx.doi.org/10.1016/j.omtn.2023.09.005
Descripción
Sumario: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(tox) 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.

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