G-quadruplex structures are key regulators of mammalian spermatogenesis

This BioProject investigates G-quadruplex (G4) structures in mouse spermatogenesis using CUT&Tag sequencing. Testes from adult male C57BL/6 mice were decapsulated and digested in Krebs buffer with 0.25% trypsin, 100 ug/ml DNase I, and 250 ug/ml collagenase I at 37 C for 21 min. Germ cells were purified via centrifugation, washing, and filtration, then separated by STA-PUT velocity sedimentation with a 2-4% BSA gradient over 3 hours. Fractions were collected, stained with Hoechst 33342 for nuclear morphology identification, and pooled into spermatogonia (SPG), spermatocytes (SPC), round spermatids (RS), and late spermatids (LS). CUT&Tag was performed on these fractions to profile G4 sites. Additionally, PDS-treated mice underwent STA-PUT to isolate SPC for targeted G4 CUT&Tag.