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

Genes to Cells : Devoted to Molecular & Cellular Mechanisms
Toshitsugu FujitaHodaka Fujii

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.

References

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Feb 6, 2016·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Toshitsugu FujitaHodaka Fujii
Jul 29, 2016·Scientific Reports·Toshitsugu FujitaHodaka Fujii
May 6, 2017·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Toshitsugu FujitaHodaka Fujii
Jun 7, 2017·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Toshitsugu FujitaHodaka Fujii

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Citations

May 13, 2020·Biology Methods and Protocols·Toshitsugu Fujita, Hodaka Fujii
May 28, 2019·Trends in Genetics : TIG·J Antonio GomezDavid J Segal
Apr 21, 2021·The CRISPR Journal·Hirotaka FujitaHodaka Fujii
May 13, 2021·Journal of Radiation Research·Kento MiuraOsamu Kaminuma
Aug 20, 2021·Biology Methods and Protocols·Miyuki YunoHodaka Fujii

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