Identification of proteins associated with an IFNγ-responsive promoter by a retroviral expression system for enChIP using CRISPR.

ABSTRACT: Isolation of specific genomic regions retaining molecular interactions is essential for comprehensive identification of molecules associated with the genomic regions. Recently, we developed the engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) technology for purification of specific genomic regions. Here, we developed a retroviral expression system for enChIP using CRISPR. We showed that the target genomic locus can be purified with high efficiency by using this system. We also showed that contamination of potential off-target sites is negligible by using this system if the guide RNA (gRNA) for the target site has a sufficiently long unique sequence in its seed sequence. enChIP combined with stable isotope labeling using amino acids in cell culture (SILAC) ana

SUBMITTER: Fujita T 

PROVIDER: S-EPMC4106880 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature