TICRR/TRESLIN and MTBP Cut&Run in G1 synchronized HCT 116 cells

The selection of replication origins plays a crucial role in eukaryotic DNA replication. However, the mechanisms of origin selection in humans are not yet well-defined. Recent work in yeast has shown that the recruitment of Sld3-Sld7 to origins during G1 is involved in origin selection. In humans, TICRR/TRESLIN and MTBP are the orthologs of Sld3 and Sld7, respectively. In this study, we have mapped the genomic binding locations of TICRR/TRESLIN and MTBP in a population of G1 synchronized cells. We utilized HCT 116 cell lines, where all TICRR/TRESLIN or MTBP alleles were tagged with mClover, to perform Cut&Run experiments with an anti-GFP antibody that specifically binds to the tag. To obtain background signal measurements, we simultaneously conducted Cut&Run experiments on the untagged cell lines. Subsequently, the generated Cut&Run reads were aligned to the genome and used to call peaks.