Base editing the mammalian genome
Base editing the mammalian genome
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DOI：
10.1016/j.ymeth.2019.02.022
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发表时间：
2019-07-01
期刊：
影响因子：
4.8
通讯作者：
Dow, Lukas E.
中科院分区：
文献类型：
--
作者：
Schatoff, Emma M.;Zafra, Maria Paz;Dow, Lukas E.
碱基编辑是一项强大的技术，可以实现哺乳动物基因组中单个核苷酸的可编程转换。碱基编辑器由部分活性的Cas9核酸酶（Cas9(D10A)）连接到天然或合成的DNA修饰酶组成。虽然最近才被描述出来，但BE已经在基础研究和转化研究中显示出巨大的潜力，可以在各种细胞类型和模式生物中创建或修复疾病等位基因。在过去的两年中，已经描述了大量新的和经过修改的基编辑器变体，扩展了该方法的灵活性和有用性。虽然概念上很简单，但有效地实现碱基编辑需要了解这些工具的优点和局限性。在这里，我们概述了DNA碱基编辑的概念，并讨论了哺乳动物细胞优化碱基编辑系统的最新进展。此外，我们强调了设计和执行BE实验的关键技术方面，并提供了细胞系和类器官中成功碱基编辑的详细实验示例，以帮助指导这些工具在基因组修饰中的有效使用。
Base editing is a powerful technology that enables programmable conversion of single nucleotides in the mammalian genome. Base editors consist of a partially active Cas9 nuclease (Cas9(D10A)) tethered to a natural or synthetic DNA modifying enzyme. Though only recently described, BE has already shown enormous potential for basic and translational research, allowing the creation or repair of disease alleles in a variety of cell types and model organisms. In the past 2 years, a vast array of new and modified base editor variants have been described, expanding the flexibility and usefulness of the approach. Though simple in concept, effective implementation of base editing requires an understanding of the advantages and limitations of each of these tools. Here, we provide an overview of the concepts of DNA base editing, and discuss the recent progress toward the development of optimized base editing systems for mammalian cells. In addition, we highlight key technical aspects of designing and executing BE experiments, and provide detailed experimental examples of successful base editing in cell lines and organoids to help guide the effective use of these tools for genome modification.