Engineering RsDddA as mitochondrial base editor with wide target compatibility and enhanced activity.
Engineering RsDddA as mitochondrial base editor with wide target compatibility and enhanced activity.
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DOI：
10.1016/j.omtn.2023.09.005
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发表时间：
2023-12-12
期刊：
影响因子：
--
通讯作者：
Lou, Xin
中科院分区：
文献类型：
--
作者：
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
双链DNA特异性胞苷脱氨酶（DddA）碱基编辑器在生物医学研究，医学和生物技术中的应用前景广阔。对间隔区域的严格序列偏好对DddA编辑器充分发挥其潜力提出了挑战。为了克服这种序列背景约束，我们分析了蛋白质数据集，并从瘤胃球菌AF 17 -6（RsDddA）中鉴定出一种新的DddAtox同系物。我们设计了RsDddA用于哺乳动物细胞系中的线粒体碱基编辑，并证明了RsDddA衍生的胞嘧啶碱基编辑器（RsDdCBE）提供了拓宽的NC序列相容性并表现出强大的编辑效率。此外，我们的研究结果表明，由RsDdCBE引起的线粒体全基因组脱靶编辑的平均频率与典型的DdCBE及其变体相当。Lou及其同事发现，来自Ruminococcus sp. AF 17 -6（RsDddA）的DddAtox同系物可以被改造成胞嘧啶碱基编辑器，并可以在哺乳动物细胞中提供更广泛的NC序列兼容性和强大的编辑效率。
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. Lou and colleagues found DddAtox homolog from Ruminococcus sp. AF17-6 (RsDddA) could be engineered into cytosine base editors and could offer a broadened NC sequence compatibility and robust editing efficiency in mammalian cells.
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