Development of RNA dependent protein switches and application to neuroscience.

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K22441
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 44:Biology at cellular to organismal levels, and related fields
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Shitamukai Atsunori 国立研究開発法人理化学研究所, 生命機能科学研究センター, 専門職研究員 (00442971)
• Project Period (FY): 2019-06-28 – 2022-03-31
• Keywords: 分子スイッチ / Cas13 / RNA

Outline of Final Research Achievements

To develop a molecular switch utilizing the specific RNA-binding activity of Cas13 protein and target crRNA, we investigated the effect of the following aspects: Cas13 protein, intein, linker property, number of and length of crRNA, and localization signal for controlling the non-specific interaction. As a result, we succeeded in creating two switch molecules with different activities based on the different properties of an intein in the cultured cell-based assay system. There are still problems to be overcome in terms of practical use, because of the high background and low activity. To overcome both, it is important to select highly active crRNA and study localization control domains to lower the background, and we will continue our research and aim to demonstrate it in vivo.

Free Research Field

神経科学

Academic Significance and Societal Importance of the Research Achievements

この研究の成果は、新しい原理に基づいた細胞認識ツールの開発において重要な基礎知見をもたらすともに、問題点も明確にした。本研究課題で作成した分子スイッチは、他のRNAをベースにした認識ツールや分割型のGFPなどとのコンビネーションが容易であり、今後、RNAをターゲットとする編集技術においての要素として重要であると考えられる。