ChIP-Seq mapping of the genome-wide distribution of FLAG-H2AK5acK9ac in genetic code expanded HeLa cells
To test the deposition of FLAG-H2AK5acK9ac mark across the genome, ChIP experiments coupled high-throughput sequencing were performed using antibodies against FLAG, H2AK5ac and H2AK9ac. Keywords: Genome binding/occupancy profiling by high throughput sequencing Overall design: An orthogonal genetic code-expanded system developed by Dr. Tao Liu and his colleagues (ACS Synth Biol 9, 2723-2736. 10.1021/acssynbio.0c00257) was used to incorporate acetyl-lysine into H2AK5K9 in living cells by expressing AcKRS-tRNAPyl and FLAG-H2AK5UAGK9UAG. ChIP-seq experiments were performed with antibodies against FLAG using cells cultured in the absence or presence of acetyl-lysine (AcK). This genetic code expanded approach was then applied in W833R-edited HeLa cells co-depleted of TIP60 and p300. Soluble chromatins extracted from these cells followed by ChIP-seq using antibodies against H2AK5ac and H2AK9ac were then performed.