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Title: In vivo CRISPR screening directly targeting testicular cells
Authors: Noguchi, Yuki
Onodera, Yasuhito
Miyamoto, Tatsuo
Maruoka, Masahiro  kyouindb  KAKEN_id  orcid (unconfirmed)
Kosako, Hidetaka
Suzuki, Jun
Author's alias: 野口, 勇貴
小野寺, 康仁
宮本, 達雄
圓岡, 真宏
小迫, 英尊
鈴木, 淳
Keywords: CRISPR screening
in vivo genome-wide screening
Issue Date: 13-Mar-2024
Publisher: Elsevier BV
Journal title: Cell Genomics
Volume: 4
Issue: 3
Thesis number: 100510
Abstract: CRISPR-Cas9 short guide RNA (sgRNA) library screening is a powerful approach to understand the molecular mechanisms of biological phenomena. However, its in vivo application is currently limited. Here, we developed our previously established in vitro revival screening method into an in vivo one to identify factors involved in spermatogenesis integrity by utilizing sperm capacitation as an indicator. By introducing an sgRNA library into testicular cells, we successfully pinpointed the retinal degeneration 3 (Rd3) gene as a significant factor in spermatogenesis. Single-cell RNA sequencing (scRNA-seq) analysis highlighted the high expression of Rd3 in round spermatids, and proteomics analysis indicated that Rd3 interacts with mitochondria. To search for cell-type-specific signaling pathways based on scRNA-seq and proteomics analyses, we developed a computational tool, Hub-Explorer. Through this, we discovered that Rd3 modulates oxidative stress by regulating mitochondrial distribution upon ciliogenesis induction. Collectively, our screening system provides a valuable in vivo approach to decipher molecular mechanisms in biological processes.
Description: マウス精巣を用いた個体内遺伝子スクリーニング系の開発. 京都大学プレスリリース. 2024-03-06.
Toward understanding sperm quality. 京都大学プレスリリース. 2024-03-06.
Rights: © 2024 The Author(s).
This is an open access article under the CC BY-NC-ND license.
DOI(Published Version): 10.1016/j.xgen.2024.100510
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