Establishments of the new forward genetic method using homologues recombination in human iPS cells

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K15420
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Dermatology
• Research Institution: Osaka University
• Principal Investigator: Yoshimura Yasuhide 大阪大学, 医学系研究科, 助教 (60263307)
• Co-Investigator(Renkei-kenkyūsha): Igawa Ken 東京医科歯科大学, 医歯学総合病院, 准教授 (00372441)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: iPS細胞 / revertant mosaicism / 遺伝性皮膚疾患 / CRISPR/Cas9 / 相同組換え

Outline of Final Research Achievements

In order to establish a forward genetic system in human iPS cells to identify the unknown mutation related skin disease, we introduced loss of heterogeneity (LOH) by inducing chromosomal crossing over, under Bloom syndrome gene (BLM) deficient condition. The Tet-off system was introduced into the BLM gene Under the control of BLM gene expression, we successfully identified clones with occurring chromosomal crossing over. By checking the SNP, we confirmed homogeneity. This indicated that this clone have occurred homologous recombination on these region. Using this system, we are trying to identify the unknown mutation in some iPS cell lines related skin disease.

Free Research Field

幹細胞生物学、遺伝学