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在 CRISPR/dCas9 敲入小鼠中对基因组区域进行实时成像和跟踪。

Live imaging and tracking of genome regions in CRISPR/dCas9 knock-in mice.

机构信息

Graduate Program, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China.

National Institute of Biological Sciences, Beijing, 102206, China.

出版信息

Genome Biol. 2018 Nov 8;19(1):192. doi: 10.1186/s13059-018-1530-1.

Abstract

CRISPR/dCas9 is a versatile tool that can be used to recruit various effectors and fluorescent molecules to defined genome regions where it can modulate genetic and epigenetic markers, or track the chromatin dynamics in live cells. In vivo applications of CRISPR/dCas9 in animals have been challenged by delivery issues. We generate and characterize a mouse strain with dCas9-EGFP ubiquitously expressed in various tissues. Studying telomere dynamics in these animals reveals surprising results different from those observed in cultured cell lines. The CRISPR/dCas9 knock-in mice provide an important and versatile tool to mechanistically study genome functions in live animals.

摘要

CRISPR/dCas9 是一种通用工具,可用于招募各种效应子和荧光分子到定义的基因组区域,从而调节遗传和表观遗传标记,或跟踪活细胞中的染色质动力学。CRISPR/dCas9 在动物体内的应用受到递送问题的挑战。我们生成并表征了一种在各种组织中普遍表达 dCas9-EGFP 的小鼠品系。在这些动物中研究端粒动力学揭示了与在培养细胞系中观察到的不同的令人惊讶的结果。CRISPR/dCas9 基因敲入小鼠为在活体动物中从机制上研究基因组功能提供了一个重要而通用的工具。

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