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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-05-01
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| Series: | Stem Cell Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506120301008 |
| _version_ | 1818127839417860096 |
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| author | Anna Karolina Blöchinger Johanna Siehler Katharina Wißmiller Alireza Shahryari Ingo Burtscher Heiko Lickert |
| author_facet | Anna Karolina Blöchinger Johanna Siehler Katharina Wißmiller Alireza Shahryari Ingo Burtscher Heiko Lickert |
| author_sort | Anna Karolina Blöchinger |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-11T07:23:44Z |
| format | Article |
| id | doaj.art-b3f8a7273c8145ada236b664f5a550a3 |
| institution | Directory Open Access Journal |
| issn | 1873-5061 |
| language | English |
| last_indexed | 2024-12-11T07:23:44Z |
| publishDate | 2020-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-b3f8a7273c8145ada236b664f5a550a32022-12-22T01:16:02ZengElsevierStem Cell Research1873-50612020-05-0145Generation of an INSULIN-H2B-Cherry reporter human iPSC lineAnna Karolina Blöchinger0Johanna Siehler1Katharina Wißmiller2Alireza Shahryari3Ingo Burtscher4Heiko Lickert5Institute of Diabetes and Regeneration Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Institute of Stem Cell Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Technische Universität München, Ismaninger Straße 22, 81675 München, GermanyInstitute of Diabetes and Regeneration Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Institute of Stem Cell Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Technische Universität München, Ismaninger Straße 22, 81675 München, GermanyInstitute of Diabetes and Regeneration Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Institute of Stem Cell Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Technische Universität München, Ismaninger Straße 22, 81675 München, GermanyInstitute of Diabetes and Regeneration Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Institute of Stem Cell Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Technische Universität München, Ismaninger Straße 22, 81675 München, GermanyInstitute of Diabetes and Regeneration Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Institute of Stem Cell Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; German Center for Diabetes Research (DZD), 85764 Neuherberg, GermanyInstitute of Diabetes and Regeneration Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Institute of Stem Cell Research, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Technische Universität München, Ismaninger Straße 22, 81675 München, Germany; German Center for Diabetes Research (DZD), 85764 Neuherberg, Germany; Corresponding author.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.http://www.sciencedirect.com/science/article/pii/S1873506120301008 |
| spellingShingle | Anna Karolina Blöchinger Johanna Siehler Katharina Wißmiller Alireza Shahryari Ingo Burtscher Heiko Lickert Generation of an INSULIN-H2B-Cherry reporter human iPSC line Stem Cell Research |
| title | Generation of an INSULIN-H2B-Cherry reporter human iPSC line |
| title_full | Generation of an INSULIN-H2B-Cherry reporter human iPSC line |
| title_fullStr | Generation of an INSULIN-H2B-Cherry reporter human iPSC line |
| title_full_unstemmed | Generation of an INSULIN-H2B-Cherry reporter human iPSC line |
| title_short | Generation of an INSULIN-H2B-Cherry reporter human iPSC line |
| title_sort | generation of an insulin h2b cherry reporter human ipsc line |
| url | http://www.sciencedirect.com/science/article/pii/S1873506120301008 |
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