Use of genome-wide RNAi screening to identify modes of action of bioactive compounds

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K12759
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Chemical biology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Matsumoto Ken 国立研究開発法人理化学研究所, 吉田化学遺伝学研究室, 専任研究員 (60222311)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: 化合物 / RNAi / スクリーニング

Outline of Final Research Achievements

Genome-wide RNA interference (RNAi) with pooled and barcoded short-hairpin RNA (shRNA) libraries provides a powerful tool for identifying cellular components that are relevant to the modes/mechanisms of action of bioactive compounds. We used multiplex barcode sequencing technology which enables parallel analysis of multiple samples. An shRNA library screen with this system revealed that downregulation of ATP1A1, a catalytic subunit of Na+/K+ ATPase, conferred significant sensitivity to aurilide B, a natural marine product that induces mitochondria-mediated apoptosis. Combined treatment with ouabain which inhibits Na+/K+ ATPase by targeting catalytic subunits potentiated sensitivity to aurilide B.

Free Research Field

分子生物学