RT Journal Article SR Electronic T1 Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis JF bioRxiv FD Cold Spring Harbor Laboratory SP 221820 DO 10.1101/221820 A1 Toshitsugu Fujita A1 Fusako Kitaura A1 Asami Oji A1 Naoki Tanigawa A1 Miyuki Yuno A1 Masahito Ikawa A1 Ichiro Taniuchi A1 Hodaka Fujii YR 2018 UL http://biorxiv.org/content/early/2018/01/18/221820.abstract AB We developed the engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) technology to isolate specific genomic regions while retaining their molecular interactions. In enChIP, the locus of interest is tagged with an engineered DNA-binding molecule, such as a modified form of the clustered regularly interspaced short palindromic repeats (CRISPR) system containing a guide RNA (gRNA) and a catalytically inactive form of Cas9 (dCas9). The locus is then affinity-purified to enable identification of associated molecules. In this study, we generated transgenic mice expressing 3xFLAG-tagged Streptococcus pyogenes dCas9 (3xFLAG-dCas9) and retrovirally transduced gRNA into primary CD4+ T cells from these mice for enChIP. Using this approach, we achieved high yields of enChIP at the targeted genomic region. Our novel transgenic mouse lines provide a valuable tool for enChIP analysis in primary mouse cells.enChIPengineered DNA-binding molecule-mediated chromatin immunoprecipitationCRISPRclustered regularly interspaced short palindromic repeatsdCas9catalytically inactive form of Cas9gRNAguide RNAGFPgreen fluorescent proteinMSmass spectrometryNGSnext-generation sequencingTgtransgenic