Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies
Summary: A major challenge in using human pluripotent stem cells (hPSCs) in therapy is the risk of teratoma formation due to contaminating undifferentiated stem cells. We used CRISPR-Cas9 for in-frame insertion of a suicide gene, iC9, into the endogenous SOX2 locus in human embryonic stem cell (ESC)...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2019-12-01
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| Series: | iScience |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004219304894 |
| _version_ | 1831751308588089344 |
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| author | Youjun Wu Tammy Chang Yan Long He Huang Fouad Kandeel Jiing-Kuan Yee |
| author_facet | Youjun Wu Tammy Chang Yan Long He Huang Fouad Kandeel Jiing-Kuan Yee |
| author_sort | Youjun Wu |
| collection | DOAJ |
| description | Summary: A major challenge in using human pluripotent stem cells (hPSCs) in therapy is the risk of teratoma formation due to contaminating undifferentiated stem cells. We used CRISPR-Cas9 for in-frame insertion of a suicide gene, iC9, into the endogenous SOX2 locus in human embryonic stem cell (ESC) line H1 for specific eradication of undifferentiated cells without affecting differentiated cells. This locus was chosen over NANOG and OCT4, two other well-characterized stem cell loci, due to significantly reduced off-target effect. We showed that undifferentiated H1-iC9 cells were induced to apoptosis by iC9 inducer AP1903, whereas differentiated cell lineages including hematopoietic cells, neurons, and islet beta-like cells were not affected. We also showed that AP1903 selectively removed undifferentiated H1-iC9 cells from a mixed cell population. This strategy therefore provides a layer of safety control before transplantation of a stem-cell-derived product in therapy. : Cellular Therapy; Techniques in Genetics; Stem Cells Research Subject Areas: Cellular Therapy, Techniques in Genetics, Stem Cells Research |
| first_indexed | 2024-12-21T22:42:40Z |
| format | Article |
| id | doaj.art-c1325e2cafec4dfc9b702089adcfe458 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-12-21T22:42:40Z |
| publishDate | 2019-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-c1325e2cafec4dfc9b702089adcfe4582022-12-21T18:47:47ZengElsevieriScience2589-00422019-12-0122409422Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based TherapiesYoujun Wu0Tammy Chang1Yan Long2He Huang3Fouad Kandeel4Jiing-Kuan Yee5Departments of Translational Research & Cellular Therapeutics, Beckman Research Institute of City of Hope, Duarte, CA 91010, USADepartments of Translational Research & Cellular Therapeutics, Beckman Research Institute of City of Hope, Duarte, CA 91010, USADepartments of Translational Research & Cellular Therapeutics, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA; Bone Marrow Transplantation Center, The First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang 310027, ChinaBone Marrow Transplantation Center, The First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang 310027, ChinaDepartments of Translational Research & Cellular Therapeutics, Beckman Research Institute of City of Hope, Duarte, CA 91010, USADepartments of Translational Research & Cellular Therapeutics, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA; Corresponding authorSummary: A major challenge in using human pluripotent stem cells (hPSCs) in therapy is the risk of teratoma formation due to contaminating undifferentiated stem cells. We used CRISPR-Cas9 for in-frame insertion of a suicide gene, iC9, into the endogenous SOX2 locus in human embryonic stem cell (ESC) line H1 for specific eradication of undifferentiated cells without affecting differentiated cells. This locus was chosen over NANOG and OCT4, two other well-characterized stem cell loci, due to significantly reduced off-target effect. We showed that undifferentiated H1-iC9 cells were induced to apoptosis by iC9 inducer AP1903, whereas differentiated cell lineages including hematopoietic cells, neurons, and islet beta-like cells were not affected. We also showed that AP1903 selectively removed undifferentiated H1-iC9 cells from a mixed cell population. This strategy therefore provides a layer of safety control before transplantation of a stem-cell-derived product in therapy. : Cellular Therapy; Techniques in Genetics; Stem Cells Research Subject Areas: Cellular Therapy, Techniques in Genetics, Stem Cells Researchhttp://www.sciencedirect.com/science/article/pii/S2589004219304894 |
| spellingShingle | Youjun Wu Tammy Chang Yan Long He Huang Fouad Kandeel Jiing-Kuan Yee Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies iScience |
| title | Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies |
| title_full | Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies |
| title_fullStr | Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies |
| title_full_unstemmed | Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies |
| title_short | Using Gene Editing to Establish a Safeguard System for Pluripotent Stem-Cell-Based Therapies |
| title_sort | using gene editing to establish a safeguard system for pluripotent stem cell based therapies |
| url | http://www.sciencedirect.com/science/article/pii/S2589004219304894 |
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