ChIP-seq of MOZ (Flag-V5-BIO tagged) in mouse mouse craniofacial development

Carboxy-terminally tagged MOZ (Flag-V5-BIO tagged) was detected by ChIP-seq using anti-V5 antibody (Sigma, A7345) to precipitate chromatin associated with MOZ Overall design: A Flag-V5-BIO triple tag was added in frame to the carboxy-terminus of the Moz locus by BAC by recombineering in E. coli. The construct was used to generate homologously recombined mouse embryonic stem cells, chimeras and mice that carry the Flag-V5-BIO tag in the Moz locus. Mice homozygous for a null mutation of Moz die between embryonic day 14.5 (E14.5) and birth. Mice homozygous fo the Moz-Flag-V5-BIO allele are viable, healthy and fertile showing that the MOZ-Flag-V5-BIO protein functions normally. The first the second pharyngeal arches of E10.5 embryos homozygous for the Moz-Flag-V5-BIO allele as well as wild-type control pharyngeal arches (negative control) were used for anti-V5 chromatin immunoprecipitation to detect the distribution of tagged MOZ in the mouse genome.