Generation of an INSULIN-H2B-Cherry reporter human iPSC line
Differentiating human induced pluripotent stem cells (hiPSCs) into insulin (INS)-producing β-like cells has potential for diabetes research and therapy. Here, we generated a heterozygous fluorescent hiPSC reporter, labeling INS-producing β-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 β-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 β-like cells. Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.
|Authors||Blöchinger AK, Siehler J, Wißmiller K, Shahryari A, Burtscher I, Lickert H|
|Journal||Stem cell research|
|Publication Date||2020 Apr 22;45:101797|