Development of a new fluorescent probe for visualization of open chromatin structure in living cells

[Speaker] Daisuke Ino:1
[Co-author] Kazuho Ikeda:1, Yasushi Okada:1
1:Laboratory for Cell Polarity Regulation, QBiC, RIKEN, Japan

The chromatin structure and its dynamics in the interphase nucleus are proposed to play essential roles in the regulation of the gene expression. In particular, the open chromatin regions, where genomic DNA becomes loose and highly accessible to the gene regulatory elements, are recognized as the center for the gene expression. Thus, if the open chromatin structure can precisely be captured, we will further be able to approach the regulatory mechanisms of gene expression. So far, open chromatin regions have been identified by next-generation sequencing-based assays such as DNase-seq, FAIRE-seq, and ATAC-seq, but it is very difficult for these biochemical techniques to analyze the dynamics of the open chromatin in living cells. Therefore, there is a pressing need for the direct visualization tools of the open chromatin structure in living cells. In this study, we have developed ChrocodiLE, a new protein-based probe that selectively binds to open chromatic region. When expressed as a fusion protein with fluorescent proteins, ChrocodiLE showed similar localization patterns to euchromatic markers, and did not colocalize with heterochromatic markers. In this poster, we will present the characterizations of ChrocodiLE and discuss its potential applications in live cell imaging experiments.
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