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工程 rsDddA作为线粒体基编辑器,具有广泛的目标兼容性和增强的活动

Kai Cheng¹, Cao Li¹, Jiachuan Jin²

¹State 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.

Molecular therapy. Nucleic acids

|September 25, 2023

概括

研究人员设计了一种新的DddA同类物 (RsDddA) 用于线粒体基编辑. 这种RsDddA衍生的细胞因子基编辑器 (RsDdCBE) 显示了改进的序列兼容性和强大的效率,具有可比的目标外效应.

科学领域:

分子生物学分子生物学
遗传学遗传学是一个
生物技术是生物技术.

背景情况:

双链DNA特定的cytidine deaminase (DddA) 基编辑器是生物医学研究,医学和生物技术中宝贵的工具.
当前DDDA编辑器的一个显著限制是它们的严格序列偏好,阻碍了它们更广泛的应用.
克服序列-上下文约束对于最大限度地发挥 DddA 基础编辑器的潜力至关重要.

研究的目的:

识别和设计一种新的DddA同源,以改善序列兼容性,用于基础编辑.
开发和描述一个DddA衍生的细胞基编辑器 (RsDdCBE) 以实现高效的线粒体基编辑.
评估工程RsDdCBE在哺乳动物细胞中的编辑效率和非目标效应.

主要方法:

对蛋白质数据集的生物信息分析,以识别新的DddA同类.
来自Ruminococcus* sp. 的DddA同源的工程. *AF17-6* (RsDddA) 用于线粒体向.
在哺乳动物细胞系中演示基编辑,并分析编辑效率和非目标频率.

主要成果:

从 *Ruminococcus* sp. 来识别和设计一种新的 DddA 同类物,RsDddA,来自 *Ruminococcus* sp. *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6* *AF17-6*
源自RsDddA的细胞因子基编辑器 (RsDdCBE) 显示了扩展的NC序列兼容性.
RsDdCBE在哺乳动物细胞中表现出强大的编辑效率,其非目标编辑频率与现有的DdCBE变体相当.

结论:

工程 rsDddA 为克服 DddA 基础编辑器的序列-上下文限制提供了一个有希望的替代方案.
RsDdCBE扩展了DddA基编辑器用于线粒体基因组编辑应用程序的实用性.
开发的RsDdCBE为生物医学研究,医学和生物技术提供了一个强大的工具,具有更好的特异性.

关键词:

DdCBE DdCBE 在线阅读 MT:RNA/DNA编辑编辑基础编辑基础编辑线粒体疾病是线粒体疾病.在 mtDNA 工程方面,目标兼容性目标兼容性

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