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进化采矿和TnpB核酶的功能特征识别有效的微型基因组编辑器

Guanghai Xiang^1,2,3, Yuanqing Li^4,5,6, Jing Sun^7,8,6

¹State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China. xiangguanghai@ioz.ac.cn.

Nature biotechnology

|June 29, 2023

概括

转子子编码的TnpB蛋白质是紧的RNA引导的DNA编辑器. 研究人员发现了新的TnpB变异,包括在人体细胞中活跃的小型,高效的基因组编辑器,突出了它们对未来基因编辑应用的潜力.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
生物技术是生物技术.

背景情况:

从转子体 (IS200/IS605) 衍生出来的TnpB蛋白质作为RNA引导的DNA内核酶起作用.
它们代表了Cas12核酶的进化前体,并具有作为紧的基因组编辑工具的潜力.

研究的目的:

探索TnpB蛋白质的进化多样性.
识别和描述具有基因组编辑能力的新型TnpB变异,特别是在人类细胞中.
建立一个框架,用于在 prokaryotic 基因组中注释 TnpB 系统.

主要方法:

从64个注释的IS605成员中选TnpB蛋白质.
在大肠杆菌和人类细胞系中进行活动测试.
为TnpB系统注释开发一个计算框架.
转子子关联基因 (TAM) 和右端元素RNA (reRNA) 的表征.

主要成果:

在大肠杆菌中确定了25种活性TnpB蛋白,其中3种在人体细胞中表现出活性.
开发了从基因组序列直接预测TAM和reRNA的方法.
发现ISAam1和ISYmu1TnpBs,它们在人类细胞中表现出强大的基因组编辑.
这些新型的TnpB比SaCas9小得多,同时保持可比的编辑效率.

结论:

巨大的TnpB蛋白质的多样性为发现新的基因组编辑器提供了丰富的资源.
ISAam1和ISYmu1代表了有希望的,紧的RNA引导的基因组编辑工具,用于人类细胞中的应用.
开发的注释框架有助于识别 prokaryotic 基因组中的 TnpB 系统.

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