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Generation of an INSULIN-H2B-Cherry reporter human iPSC line.
Stem Cell Res. 45:101797 (2020)
Differentiating human induced pluripotent stem cells (hiPSCs) into insulin (INS)-producing beta-like cells has potential for diabetes research and therapy. Here, we generated a heterozygous fluorescent hiPSC reporter, labeling INS-producing beta-like cells. We used CRISPR/Cas9 technology to knock-in a T2A-H2B-Cherry cassette to replace the translational INS stop codon, enabling co-transcription and T2A-peptide mediated co-translational cleavage of INS-T2A and H2B-Cherry. The hiPSC-INS-T2A-H2B-Cherry reporter cells were pluripotent and showed multi-lineage differentiation potential. Cells expressing the beta-cell specific hormone INS are identified by nuclear localized H2B-Cherry reporter upon pancreatic endocrine differentiation. Thus, the generated reporter hiPSCs enable live identification of INS hormone-producing beta-like cells.
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Publication type
Article: Journal article
Document type
Scientific Article
ISSN (print) / ISBN
1873-5061
e-ISSN
1876-7753
Journal
Stem Cell Research
Quellenangaben
Volume: 45,
Article Number: 101797
Publisher
Elsevier
Publishing Place
Radarweg 29, 1043 Nx Amsterdam, Netherlands
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Reviewing status
Peer reviewed