Stem Cell Research (Oct 2019)

Generation of a NESTIN-EGFP reporter human induced pluripotent stem cell line, KSCBi005-A-1, using CRISPR/Cas9 nuclease

  • Youngsun Lee,
  • Hye Young Choi,
  • Ara Kwon,
  • Hyeyeon Park,
  • Mihyun Park,
  • Yong-Ou Kim,
  • Sungwook Kwak,
  • Soo Kyung Koo

Journal volume & issue
Vol. 40

Abstract

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NESTIN, an intermediate filament, is a neuroectodermal marker involved in induced pluripotent stem cell (iPSC) differentiation toward neural lineages. Here, we introduced an EGFP reporter into the C-terminus of NESTIN in KSCBi005-A hiPSCs through homologous recombination using CRISPR/Cas9 nuclease. The successfully edited line was confirmed by sequencing and had a normal karyotype. It expressed EGFP upon induction of neural differentiation and exhibited potential for differentiation into three germ layers. KSCBi005-A-1 cells could be used to monitor the expression of NESTIN in differentiated cell types. This cell line is available at the National Stem Cell Bank, Korea National Institute of Health.