| First Author | Fujita T | Year | 2018 |
| Journal | Genes Cells | Volume | 23 |
| Issue | 4 | Pages | 318-325 |
| PubMed ID | 29480524 | Mgi Jnum | J:258204 |
| Mgi Id | MGI:6144342 | Doi | 10.1111/gtc.12573 |
| Citation | Fujita T, et al. (2018) Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis. Genes Cells 23(4):318-325 |
| abstractText | 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. |
