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

引用

页数：33

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