Study for establishing a method to optically and cell-specifically manipulate single locus gene expression

2018 Fiscal Year Final Research Report

• Project/Area Number: 15K14575
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Genetics/Chromosome dynamics
• Research Institution: Shiga University of Medical Science (2016-2018); Kyoto University (2015)
• Principal Investigator: Sugi Takuma 滋賀医科大学, 神経難病研究センター, 助教 (70571305)
• Co-Investigator(Kenkyū-buntansha): 伊藤 浩史 九州大学, 芸術工学研究院, 准教授 (20512627)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Keywords: 線虫 / ゲノム編集 / 光遺伝学 / 記憶・学習 / 行動

Outline of Final Research Achievements

In this study, I established a technique, in which a gene expression of a single locus in C. elegans can be modified by genome editing technique in a cell-specific manner. I first expressed VP64 transcriptional activator in several neurons (~10 neurons) of C. elegans. This expression induces glr-1 gene in several neurons, but there were a few neurons on which VP64 has no effect. Therefore, I next tried to directly manipulate the epigenome using TALE-epigenetic factor. In this experiment, I succeeded in inducing the glr-1 expression in the neurons in which TALE-VP64 could not induce its expression. Overall, I proved that these techniques are useful to artificially induce gene expression and epigenome editing of a single locus in a cell-specific manner. Furthermore, I also insist that the techniques are also important to understand the robustness of a single locus. I am now trying to drive the gene expression and epigenome editing in a cell-specific manner by an optical manipulation.

Free Research Field

分子遺伝学

Academic Significance and Societal Importance of the Research Achievements

本手法が単純な遺伝子発現量の操作方法としてだけではなく、特定の神経細胞のクロマチン構造の「ロバストネス」を調べる方法としても有望である可能性がある. また現在, in vivoで光刺激によるエピゲノム操作を行えるように技術拡張しており、将来的に本研究により、C. elegansの記憶・学習機構を人為的に操作することにより、記憶形成に必須のエピゲノムを同定することが期待できる.