Reference - Detail
RRC ID | 81948 |
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Author | Noguchi Y, Maruoka M, Suzuki J. |
Title | Protocol for in vivo CRISPR screening targeting murine testicular cells. |
Journal | STAR Protoc |
Abstract |
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. |
Volume | 5(3) |
Pages | 103306 |
Published | 2024-9-20 |
DOI | 10.1016/j.xpro.2024.103306 |
PII | S2666-1667(24)00471-4 |
PMID | 39269899 |
PMC | PMC11416641 |
MeSH | Animals CRISPR-Cas Systems* / genetics Male Mice RNA, Guide, CRISPR-Cas Systems / genetics Spermatogenesis* / genetics Testis* / cytology Testis* / metabolism |
Resource | |
DNA material | pCAG-HIVgp (RDB04394) pCMV-VSV-G-RSV-Rev (RDB04393) pCMV-SVF-P2A-VSV-G-pRSV-REV (RDB20434) |