induction from fibroblast to keratinocyte

2014 Fiscal Year Final Research Report

• Project/Area Number: 25670500
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Dermatology
• Research Institution: Dokkyo Medical University
• Principal Investigator: SATO HITOMI 獨協医科大学, 医学部, 助教 (00648090)
• Co-Investigator(Renkei-kenkyūsha): IGAWA Ken 東京医科歯科大学, 医学部附属病院, 講師 (00372441)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: iPS細胞 / ヒト / 人工ヌクレアーゼ / 表皮角化細胞

Outline of Final Research Achievements

In order to archive our final purpose, "Establish the direct conversion system of keratinocytes from dermal fibroblasts in human", first, we tried to establish the "better" detection system for keratinocytes differentiation in human. For this purpose, keratin 14 (K14) gene that is one of major genetic markers for epidermal keratinocytes was selected and we tried to detect the on/off of this gene by visually.
With using transcription activator-like effector nucleases (TALENs), one of engineered nucleases, gene targeting for eGFP gene knock-in into the downstream of K14 gene in human iPS cells was performed. After antibiotics selection, colonies were picked up and checked transgene existences, and revealed several clones were succeeded to be targeted. Then, we differentiated these hiPS cells (K14-eGFP hiPSCs) into epidermal keratinocytes with our-established protocols and confirmed that this system works well.

Free Research Field

皮膚科学