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
State Key Laboratory of Reproductive Medicine, Women's Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing Medical University, Nanjing, Jiangsu, China.
Center for Reproductive Medicine, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Mol Ther Nucleic Acids. 2023 Sep 9;34:102028. doi: 10.1016/j.omtn.2023.09.005. eCollection 2023 Dec 12.
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 homolog from sp. (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.
双链DNA特异性胞苷脱氨酶（DddA）碱基编辑器在生物医学研究、医学和生物技术应用中具有巨大潜力。对间隔区域严格的序列偏好给DddA编辑器充分发挥其潜力带来了挑战。为了克服这种序列上下文限制，我们分析了一个蛋白质数据集，并从 种中鉴定出一种新型DddA同源物（RsDddA）。我们对RsDddA进行工程改造，用于哺乳动物细胞系中的线粒体碱基编辑，并证明源自RsDddA的胞嘧啶碱基编辑器（RsDdCBE）具有更广泛的NC序列兼容性，并表现出强大的编辑效率。此外，我们的结果表明，RsDdCBE引起的线粒体全基因组脱靶编辑的平均频率与经典DdCBE及其变体相当。
