Novel shRNA library screening to identify key factors that are important in pluripotent stem cells

Research Project

Project/Area Number	24770206
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Developmental biology
Research Institution	Kyoto University
Principal Investigator	HOTTA Akitsu 京都大学, iPS細胞研究所, 特定拠点助教 (50578002)
Project Period (FY)	2012-04-01 – 2015-03-31
Project Status	Completed (Fiscal Year 2014)
Budget Amount *help	¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000) Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000) Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywords	shRNAライブラリー / トランスポゾンベクター / 発現抑制制御 / 発現抑制
Outline of Final Research Achievements	To reveal the molecular mechanisms behind the pluripotency of ES cells and iPS cells, we constructed a piggyBac DNA transposon vector to deliver a genome-wide shRNA library into ES/iPS cells. The generated shRNA library was validated by deep sequencing, and several ten-thousand shRNAs were successfully cloned into our piggyBac vector. Our piggyBac shRNA library is a versatile tool to dissect the molecular mechanisms in pluripotent stem cells and other cell types, without the use of viral vectors.