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Stem Cell Res. 2019 Aug 27;40:101554. doi: 10.1016/j.scr.2019.101554. [Epub ahead of print]

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

Author information

1
Division of Intractable Diseases, National Center for Stem cell and Regenerative Medicine, Korea National Institute of Health, Chungcheongbuk-do, South Korea.
2
Division of Intractable Diseases, National Center for Stem cell and Regenerative Medicine, Korea National Institute of Health, Chungcheongbuk-do, South Korea. Electronic address: kswin3@korea.kr.
3
Division of Intractable Diseases, National Center for Stem cell and Regenerative Medicine, Korea National Institute of Health, Chungcheongbuk-do, South Korea. Electronic address: skkoo@nih.go.kr.

Abstract

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.

PMID:
31499408
DOI:
10.1016/j.scr.2019.101554

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