biostudies-literatureChen QNIEHS NIH HHSZ01ES102745NIH HHSdev204557https://www.ebi.ac.uk/biostudies/studies/S-EPMC12401510biostudies-literatureUnknown152(15)The foundation of spermatogenesis and lifelong fertility is provided by spermatogonial stem cells (SSCs). SSCs divide asymmetrically to either self-renew or produce undifferentiated progenitors. However, regulatory mechanisms governing SSC maintenance are poorly understood. Here, we show that the CCR4-NOT mRNA deadenylase complex subunit CNOT3 is essential for sustaining spermatogonial populations in mice. Its deletion in adult germ cells resulted in germ cell loss and infertility, and its deletion in spermatogonia in the developing testis resulted in SSC depletion and compromised spermatogenesis. Consistent with the in vivo results, deletion of Cnot3 in cultured SSCs caused a reduction in cell proliferation and viability, and downregulation of SSC markers. Mechanistically, Cnot3 deletion Development (Cambridge, England)Cnot3 is required for male germ cell development and spermatogonial stem cell maintenance.PMC12401510R21HD105963R01HD09003Geyer CBChen QKirsanov OMalki SZheng XLackford BLHu GBennett BXu XWang JfalseCnot3 is required for male germ cell development and spermatogonial stem cell maintenance.The foundation of spermatogenesis and lifelong fertility is provided by spermatogonial stem cells (SSCs). SSCs divide asymmetrically to either self-renew or produce undifferentiated progenitors. However, regulatory mechanisms governing SSC maintenance are poorly understood. Here, we show that the CCR4-NOT mRNA deadenylase complex subunit CNOT3 is essential for sustaining spermatogonial populations in mice. Its deletion in adult germ cells resulted in germ cell loss and infertility, and its deletion in spermatogonia in the developing testis resulted in SSC depletion and compromised spermatogenesis. Consistent with the in vivo results, deletion of Cnot3 in cultured SSCs caused a reduction in cell proliferation and viability, and downregulation of SSC markers. Mechanistically, Cnot3 deletion 2025-01-01T00:00:00Z2025 Aug2026-05-29T11:37:30.41Z2026-04-08T04:29:38.101ZS-EPMC124015104081496410.1242/dev.204557