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bioRxiv · 10.1101/221820

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

Fujita, T.·Kitaura, F.·Oji, A.·Tanigawa, N.·Yuno, M.·Ikawa, M.·Taniuchi, I.·Fujii, H.
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Abstract

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.

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Fujita, T., Kitaura, F., Oji, A., Tanigawa, N., Yuno, M., Ikawa, M., Taniuchi, I., Fujii, H.. 2017-11-18. Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis. https://doi.org/10.1101/221820

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