RRC ID 81948
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)