Generation of a TALEN-mediated, p63 knock-in in human induced pluripotent stem cells

• Main Authors: Yuki Kobayashi, Ryuhei Hayashi, Andrew J. Quantock, Kohji Nishida
• Format: Article
• Language: English
• Published: Elsevier 2017-12-01
• Series: Stem Cell Research
• Subjects: Human iPS cells; TALEN; Knock-in technology; p63; Stem cells; Epithelial stem cells; Corneal epithelium
• Online Access: http://www.sciencedirect.com/science/article/pii/S1873506117302179

The expression of p63 in surface ectodermal cells during development of the cornea, skin, oral mucosa and olfactory placodes is integral to the process of cellular self-renewal and the maintenance of the epithelial stem cell status. Here, we used TALEN technology to generate a p63 knock-in (KI) human induced pluripotent stem (hiPS) cell line in which p63 expression can be visualized via enhanced green fluorescent protein (EGFP) expression. The KI-hiPS cells maintained pluripotency and expressed the stem cell marker gene, ΔNp63α. They were also able to successfully differentiate into functional corneal epithelial cells as assessed by p63 expression in reconstructed corneal epithelium. This approach enables the tracing of p63-expressing cell lineages throughout epithelial development, and represents a promising application in the field of stem cell research.