Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis

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  • Transgenic mouse lines expressing the 3x<scp>FLAG</scp>‐<scp>dC</scp>as9 protein for enCh<scp>IP</scp> analysis

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Abstract

Fujita, T, Kitaura, F, Oji, A, et al. Transgenic mouse lines expressing the 3xFLAG‐dCas9 protein for enChIP analysis. Genes Cells. 2018; 23: 318– 325. https://doi.org/10.1111/gtc.12573

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.

Journal

  • Genes to Cells

    Genes to Cells 23 (4), 318-325, 2018-04-10

    Blackwell Publishing Ltd

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