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Category | CDBセミナー |
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Date and Time | 2016-03-15 16:00 - 17:00 |
Venue | Seminar Room A7F |
Speaker | Hodaka Fujii |
Affiliation | Chromatin Biochemistry Research Group, Combined Program on Microbiology and Immunology, Research Institute for Microbial Diseases, Osaka University |
Title | Biochemical analysis of genome functions using the locus-specific chromatin immunoprecipitation technologies: key tools to elucidate 4D Nucleome |
Poster | click here to download(PDF) |
Host | Ichiro Hiratani |
Abstract | Elucidation of molecular mechanisms of genome functions such as transcription and epigenetic regulation requires identification of components mediating the genome functions. To this end, we developed the locus-specific chromatin immunoprecipitation (locus-specific ChIP) technologies, consisting of insertional ChIP (iChIP) and engineered DNA-binding molecule-mediated ChIP (enChIP) using transcription activator-like (TAL) proteins and the clustered regularly interspaced short palindromic repeats (CRISPR) system. Locus-specific ChIP consists of locus tagging and affinity purification and can be combined with down-stream analyses such as mass spectrometry (MS), RNA sequencing (RNA-Seq), or next-generation sequencing to identify proteins, RNAs, or other genomic regions associated with the target genomic region. I will present our recent analyses of genome functions using locus-specific ChIP. We have successfully identified proteins associated with even a single copy locus of multicellular higher eukaryotes using locus-specific ChIP combined with MS. Furthermore, using enChIP-RNA-Seq, we identified RNAs associated with telomeres. To our knowledge, this is the first non-biased identification of RNAs associated with specific genomic regions. Finally, we've successfully identified interactions between genomic regions using iChIP-Seq and enChIP-Seq analyses, showing that these technologies would be key tools for elucidation of "4D Nucleome", dynamics of the three-dimensional structure of the nucleus. (This seminar is a part of the Epigenetics Seminar Series 2015) |