Generation of a NESTIN-EGFP reporter human induced pluripotent stem cell line, KSCBi005-A-1, using CRISPR/Cas9 nuclease
| Autor: | Hye Young Choi, Ara Kwon, Young-Sun Lee, Yong-Ou Kim, Hyeyeon Park, Soo Kyung Koo, Mi-Hyun Park, Sungwook Kwak |
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| Rok vydání: | 2019 |
| Předmět: |
Male
0301 basic medicine Cellular differentiation Green Fluorescent Proteins Induced Pluripotent Stem Cells Germ layer Biology Cell Line Nestin 03 medical and health sciences 0302 clinical medicine Genes Reporter Humans CRISPR Induced pluripotent stem cell lcsh:QH301-705.5 Nuclease Cell Differentiation Cell Biology General Medicine Cell biology 030104 developmental biology lcsh:Biology (General) Cell culture biology.protein CRISPR-Cas Systems Stem cell 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Stem Cell Research, Vol 40, Iss, Pp-(2019) |
| ISSN: | 1873-5061 |
| DOI: | 10.1016/j.scr.2019.101554 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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