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Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System
Published on: May 28, 2015
Generation of a PDX1-EGFP reporter human induced pluripotent stem cell line, KSCBi005-A-3, using the CRISPR/Cas9
Youngsun Lee1, Hye Young Choi1, Ara Kwon1
1Division of Intractable Diseases, National Center for Stem cell and Regenerative Medicine, Korea National Institute of Health, Chungcheongbuk-do, South Korea.
Abstract:
PDX1 plays a crucial role in the development and maintenance of β-cells and directly regulates pancreatic β-cell-specific transcription factors by binding to the insulin gene. Here, we introduced an EGFP reporter into the C-terminus of PDX1 in KSCBi005-A human induced pluripotent stem cells through homologous recombination using CRISPR/Cas9 nuclease. The cells had a normal karyotype, expressed several pluripotency markers, and maintained their differentiation potential. KSCBi005-A-3 cells can be used to monitor PDX1 expression in live cells during β-cell differentiation; the cell line has been registered at the National Stem Cell Bank, Korea National Institute of Health.
