Identification of endogenous G-quadruplexes (G4s) in human, mouse, and pig (CUT&Tag)

G-quadruplexes (G4s) are non-canonical four-stranded structures and emerge as novel genetic regulatory elements. Endogenous G4s (eG4s) can form quadruplex structures and induce double strand breaking during DNA replication, and thus contributing to genomic instability and genome evolution. Besides, eG4s are closely associated with gene expression regulation, and both their stimulative and repressive roles in the genome have been elucidated. Although functions of several specific G4s have been reported, a systematical identification and characterization of eG4s is still unconducted. Here we applied Cleavage Under Targets and Tagmentation (CUT&Tag) for mapping eG4s across human, mouse, and pig at high resolution and low background. We analyzed their genomics distribution, evolutionary conservation, dynamics, regulation, and impacts on gene expression in a single framework. CUT&Tag using G4-specific antibody, BG4 Overall design: We performed G4 CUT&Tag experiment on genomic DNA extracted from human, mouse, and pig cells in biological triplicate. HCT116, SW620, A549, HT29, 293T, HepG2, and NCM460 were from human, c2c12 and 3T3 were from mouse, and PK15 and ST were from pig. We sequenced a total of 30 samples.