Comprehensive analysis of mutant mammalian cells using barcode vector

Research Project

Project/Area Number	23650235
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Laboratory animal science
Research Institution	Osaka University
Principal Investigator	HORIE Kyoji 大阪大学, 医学系・研究科, 准教授 (30333446)
Project Period (FY)	2011
Project Status	Completed (Fiscal Year 2011)
Budget Amount *help	¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000) Fiscal Year 2011: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Keywords	発生工学 / ES細胞 / バーコード / 変異体
Research Abstract	We previously developed a method for rapid isolation of a large number of homozygous mutant mouse ES cell lines. However, we realized that phenotypic analysis of each mutant cell line is a labor-intensive process. In the current study, we have constructed a barcode vector library in which each vector contains a unique nucleotide sequence, and labeled each homozygous mutant cell line with different barcode sequence. This allowed us to evaluate the ratio of each mutant cell line among entire population of a pool of mutant cells using each barcode sequence as an index to represent the corresponding mutant cell line, leading to a rapid phenotypic analysis with pooled cells.