Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis
| Autor: | Naoki Tanigawa, Hodaka Fujii, Masahito Ikawa, Miyuki Yuno, Fusako Kitaura, Asami Oji, Toshitsugu Fujita, Ichiro Taniuchi |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
enChIP
CD4-Positive T-Lymphocytes 0301 basic medicine Genetically modified mouse Chromatin Immunoprecipitation ChIP Mice Transgenic Locus (genetics) Biology medicine.disease_cause dCas9 Mice 03 medical and health sciences CRISPR-Associated Protein 9 Genetics medicine Animals CRISPR Clustered Regularly Interspaced Short Palindromic Repeats Guide RNA Cells Cultured Cas9 Palindrome Cell Biology Cell biology transgenic mouse Mice Inbred C57BL 030104 developmental biology Gene Expression Regulation Streptococcus pyogenes Genetic Engineering Chromatin immunoprecipitation RNA Guide Kinetoplastida |
| DOI: | 10.1101/221820 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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