A TALEN-mediated, p63 knock-in application in human iPS cells
Authors
Kobayashi, Y, Hayashi, R, Quantock, Andrew and Kohji Nishida, K
Institution
Osaka University
Country
Japan
Year
2017
Abstract
The p63 is a biomarker whose expression in surface ectodermal cells during
developmental processes of the cornea, skin, oral mucosa, and olfactory
placodes and is indispensable for understanding the mechanisms of self-renewal
and maintenance of epithelial stemness. Here, we generated a p63 knock-in (KI)
human induced pluripotent stem (hiPS) cell line by using TALEN, where p63
expression was visualized via enhanced green fluorescent protein expression.
The established cells maintained pluripotency, and were found to be specifically
targeted at the stem cell marker gene, ΔNp6γα. The KI-hiPS cells successfully
differentiated into corneal epithelial cells, and the functional epithelial stem cells
through p63 expression were used to evaluate reconstruction of the corneal
epithelium.This novel approach enables the tracing of cell lineages expressing p63 from the early differentiation stage throughout epithelial development.
Therefore, our data indicated that the p63-hiPS KI cells can be used as an
effective tool extending beyond studies of epithelial cell lineages