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通过哺乳动物细胞的连续定向进化系统,进化了高精度的丁和丁基编辑器,并最大限度地减少了目标外活动

Na Zhao^1,2, Jian Zhou^3,4, Tianfu Tao¹

¹Precise Genome Engineering Center, School of Life Sciences, Guangzhou University, Guangzhou, China.

Nature communications

|September 17, 2024

概括

研究人员在哺乳动物细胞中开发了一种持续定向进化系统 (CDEM),用于基础编辑器 (BEs). 该系统使哺乳动物细胞中BEs的高效进化成为可能,提高了编辑精度并减少了非目标效应.

科学领域:

分子生物学分子生物学
基因编辑技术的技术
生物技术是生物技术.

背景情况:

持续定向进化在细菌中取得了成功,用于基编辑器 (BE) 开发.
现有的方法在哺乳动物细胞系统中面临局限性,阻碍了体内应用.

研究的目的:

在哺乳动物细胞中开发一种新的持续定向进化系统 (CDEM).
为了使基编辑器 (BEs) 在哺乳动物细胞环境中的进化,而没有尺寸限制.

主要方法:

开发CDEM系统,使用Cas9尼克酶进行全长的基础编辑器演变.
应用CDEM来发展细胞因子基编辑器BE3和腺因基编辑器 (ABEmax,ABE8e).

主要成果:

进化的cytidine deaminase变体显示了缩小编辑窗口,提高纯度,并减少了目标外活动.
进化的腺基编辑器变体显示了更改的编辑窗口和减少的目标外效应.
在哺乳动物细胞中,CDEM证明对连续基编辑器进化有效.

结论:

CDEM是一个强大而通用的平台,用于在哺乳动物系统中推进基编辑器技术.
开发的系统克服了基于细菌进化的局限性,提供了更广泛的适用性.
CDEM有助于创建更精确,更有效的基因编辑工具,用于治疗开发.

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