Protocol for in vivo CRISPR screening targeting murine testicular cells

被引：0
|
作者：
Noguchi, Yuki [1 ,2 ]
Maruoka, Masahiro [1 ]
Suzuki, Jun [1 ,2 ,3 ,4 ]
机构：
[1] Kyoto Univ, Inst Integrated Cell Mat Sci WPI iCeMS, Yoshida Honmachi,Sakyoku, Kyoto 6068501, Japan
[2] Kyoto Univ, Grad Sch Biostudies, Konoe Cho,Sakyoku, Kyoto 6068501, Japan
[3] Acad Sinica, Inst Biomed Sci, Ctr Integrated Biosyst, Taipei, Taiwan
[4] Japan Sci & Technol Agcy, CREST, Kawaguchi, Saitama 3320012, Japan
来源：
STAR PROTOCOLS | 2024年 / 5卷 / 03期
基金：
日本科学技术振兴机构;
关键词：
STEM-CELLS; MICE;
D O I：
10.1016/j.xpro.2024.103306
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
In vivo genome-wide screening elucidates tissue-specific molecular events. Here, we present a protocol for an in vivo genome-wide CRISPR-Cas9 single- guide RNA (sgRNA) library screening technique optimized for mouse testicular cells to investigate spermatogenesis. We describe steps for virus injection, sperm sorting, and primase-based whole-genome amplification. We then detail procedures for library reconstruction using a "revival screening"technique. Our approach reveals intricate spermatogenesis processes and is adaptable for diverse tissue-specific studies. For complete details on the use and execution of this protocol, please refer to Noguchi et al.1 1
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页数：33
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