Precise spatiotemporal gene expression during animal development is achieved through gene regulatory networks, in which sequence-specific transcription factors (TFs) bind to cis-regulatory elements of target genes. Although numerous cis-regulatory elements have been identified in a variety of systems, their global architecture in the gene networks that regulate animal development is not well understood. Here, we determined the structure of the core networks at the cis-regulatory level in early e ...[more]
Project description:DNA binding profiles of three maternal factors (Tcf7, Gata.a and Zic-r.a) in chordate 16-cell stage embryo. 16-cell stage ciona intestinalis embryos were collected from multiple batches and the extracted chromatin was immunoprecipitated by the antibodies which specifically recognize ciona Gata.a, Tcf7 and Zic-r.a, respectively. Overall design: Three independent chromatin immunoprecipitation assay (Gata.a, Tcf7 and Zic-r.a)
Project description:A genomic overview of in vivo binding of a transcription factor FoxA-a in the ascidian 64-cell stage embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged FoxA-a. After fixation, Chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.