Dissection of anterior mesendoderm segregation at single cell level in zebrafish

Purpose: To investigate Ripply1 binding site in zebrafish embryo Methods: HA-ripply-injected embryos were harvested at 6hpf. CUT&Tag were performed on the collected cells using Hyperactive Universal CUT&Tag Assay Kit for Illumina Pro (Vazyme, TD904) following the protocol.The products were sent for paired-end sequencing at 150 bp read length on a NovaSeq 6000 system. Results: 72,683 peaks were enriched in HA-ripply1-injected samples. And Ripply1 binding peaks were found to be enriched in the immediate upstream region of the gene body of sox17 and sox32. Conclusions: Ripply1 may repress sox17 and sox32 expression through direct binding. Zebrafish embryos injected with HA-ripply1 mRNA and were harvested at 6hpf. Anti-HA antibody was then used to extract HA-Ripply1 for CUT&Tag experiment.