Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells

Sakthi D. Moorthy; Jennifer A. Mitchell

doi:10.3791/53552

Journal

/

发育生物学

/

生成CRISPR / Cas9介导的单等位基因缺失来研究小鼠胚胎干细胞功能增强

Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells

JoVE 杂志
发育生物学

需要订阅 JoVE 才能查看此. 登录或开始免费试用。

JoVE 杂志发育生物学

Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells

生成CRISPR / Cas9介导的单等位基因缺失来研究小鼠胚胎干细胞功能增强

Please note that all translations are automatically generated. Click here for the English version.

14,055 Views

•

11:31 min

•

April 02, 2016

DOI:

10.3791/53552-v

DOI:

10.3791/53552-v

•

11:31 min

April 02, 2016

•

14052 Views

Sakthi D. Moorthy¹, Jennifer A. Mitchell¹

¹Department of Cell and Systems Biology,University of Toronto

Summary

Experimental validation of enhancer activity is best approached by loss-of-function analysis. Presented here is an efficient protocol that uses CRISPR/Cas9 mediated deletion to study allele-specific regulation of gene transcription in F1 ES cells which contain a hybrid genome (Mus musculus¹²⁹ x Mus castaneus).