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

Yuki Kobayashi, Ryuhei Hayashi, Andrew J. Quantock, Kohji Nishida
2017 Stem Cell Research  
Highlights 19  A p63 knock-in cell line in human iPS cells was generated by TALEN gene 20 editing.  Visualizing p63+ cells facilitated to trace the epithelial differentiation. 1  Isolating p63+ cells during differentiation enabled epithelial stem cell research. 2  p63 knock-in human iPS cells apply for the studies of epithelial cell lineages. 3 4 Keywords 5 Abstract 7 The p63 is a biomarker whose expression in surface ectodermal cells during 8 developmental processes of the cornea, skin,
more » ... he cornea, skin, oral mucosa, and olfactory 9 placodes and is indispensable for understanding the mechanisms of self-renewal 10 and maintenance of epithelial stemness. Here, we generated a p63 knock-in (KI) 11 human induced pluripotent stem (hiPS) cell line by using TALEN, where p63 12 expression was visualized via enhanced green fluorescent protein expression. 13 The established cells maintained pluripotency, and were found to be specifically 14 targeted at the stem cell marker gene, ΔNp6γα. The KI-hiPS cells successfully 15 differentiated into corneal epithelial cells, and the functional epithelial stem cells 16 through p63 expression were used to evaluate reconstruction of the corneal 17 epithelium. This novel approach enables the tracing of cell lineages expressing 18 3 p63 from the early differentiation stage throughout epithelial development.
doi:10.1016/j.scr.2017.10.015 pmid:29179035 fatcat:mux2wh523vgufnpd6twigackwi