Comparative RNAi Screens in Isogenic Human Stem Cells Reveal SMARCA4 as a Differential Regulator
Summary: Large-scale RNAi screens are a powerful approach to identify functions of genes in a cell-type-specific manner. For model organisms, genetically identical (isogenic) cells from different cell types are readily available, making comparative studies meaningful. However, large-scale screens in...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2019-05-01
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| Series: | Stem Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213671119300979 |
| _version_ | 1811276046747566080 |
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| author | Ceren Güneş Maciej Paszkowski-Rogacz Susann Rahmig Shahryar Khattak Aylin Camgöz Martin Wermke Andreas Dahl Martin Bornhäuser Claudia Waskow Frank Buchholz |
| author_facet | Ceren Güneş Maciej Paszkowski-Rogacz Susann Rahmig Shahryar Khattak Aylin Camgöz Martin Wermke Andreas Dahl Martin Bornhäuser Claudia Waskow Frank Buchholz |
| author_sort | Ceren Güneş |
| collection | DOAJ |
| description | Summary: Large-scale RNAi screens are a powerful approach to identify functions of genes in a cell-type-specific manner. For model organisms, genetically identical (isogenic) cells from different cell types are readily available, making comparative studies meaningful. However, large-scale screens in isogenic human primary cells remain challenging. Here, we show that RNAi screens are possible in genetically identical human stem cells, using induced pluripotent stem cells as intermediates. The screens revealed SMARCA4 (SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4) as a stemness regulator, while balancing differentiation distinctively for each cell type. SMARCA4 knockdown in hematopoietic stem and progenitor cells caused impaired self-renewal in vitro and in vivo with skewed myeloid differentiation; whereas, in neural stem cells, it impaired self-renewal while biasing differentiation toward neural lineage, through combinatorial SWI/SNF subunit assembly. Our findings pose a powerful approach for deciphering human stem cell biology and attribute distinct roles to SMARCA4 in stem cell maintenance. : Güneş et al. show that RNAi screens on genetically identical (isogenic) human stem cells can dissect epigenetic factors important for self-renewal versus differentiation. By using iPSCs as a bridging cell type, the comparative RNAi screens in HSPCs and NSCs identify SMARCA4 as an important regulator of self-renewal and differentiation with cell-type-specific functions. Keywords: RNAi, comparative functional profiling, isogenic, hematopoiesis, self-renewal, neural differentiation, SMARCA4, SWI/SNF |
| first_indexed | 2024-04-12T23:49:34Z |
| format | Article |
| id | doaj.art-c7e4e0564b58471387e6cfe367555d57 |
| institution | Directory Open Access Journal |
| issn | 2213-6711 |
| language | English |
| last_indexed | 2024-04-12T23:49:34Z |
| publishDate | 2019-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Reports |
| spelling | doaj.art-c7e4e0564b58471387e6cfe367555d572022-12-22T03:11:44ZengElsevierStem Cell Reports2213-67112019-05-0112510841098Comparative RNAi Screens in Isogenic Human Stem Cells Reveal SMARCA4 as a Differential RegulatorCeren Güneş0Maciej Paszkowski-Rogacz1Susann Rahmig2Shahryar Khattak3Aylin Camgöz4Martin Wermke5Andreas Dahl6Martin Bornhäuser7Claudia Waskow8Frank Buchholz9Medical Faculty and University Hospital Carl Gustav Carus, UCC Section Medical Systems Biology, TU Dresden, 01307 Dresden, GermanyMedical Faculty and University Hospital Carl Gustav Carus, UCC Section Medical Systems Biology, TU Dresden, 01307 Dresden, GermanyRegeneration in Hematopoiesis, Leibniz Institute on Aging – Fritz Lipmann Institute, 07745 Jena, GermanyStem Cell Engineering Facility, Biotechnology Center, TU Dresden, 01307 Dresden, GermanyMedical Faculty and University Hospital Carl Gustav Carus, UCC Section Medical Systems Biology, TU Dresden, 01307 Dresden, Germany; National Center for Tumor Diseases (NCT), University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, GermanyDepartment of Medicine I, University Hospital Carl Gustav Carus, 01307 Dresden, Germany; Medical Faculty and University Hospital Carl Gustav Carus, Early Clinical Trial Unit, 01307 Dresden, GermanyDresden Genome Center (DGC), TU Dresden, 01307 Dresden, GermanyDepartment of Medicine I, University Hospital Carl Gustav Carus, 01307 Dresden, GermanyRegeneration in Hematopoiesis, Leibniz Institute on Aging – Fritz Lipmann Institute, 07745 Jena, Germany; Department of Medicine III, Faculty of Medicine, Fetscherstrasse 74, 01307 Dresden, GermanyMedical Faculty and University Hospital Carl Gustav Carus, UCC Section Medical Systems Biology, TU Dresden, 01307 Dresden, Germany; German Cancer Research Center (DKFZ), Heidelberg and German Research Consortium (DKTK), Partner Site Dresden, 01307 Dresden, Germany; National Center for Tumor Diseases (NCT), University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany; Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany; Corresponding authorSummary: Large-scale RNAi screens are a powerful approach to identify functions of genes in a cell-type-specific manner. For model organisms, genetically identical (isogenic) cells from different cell types are readily available, making comparative studies meaningful. However, large-scale screens in isogenic human primary cells remain challenging. Here, we show that RNAi screens are possible in genetically identical human stem cells, using induced pluripotent stem cells as intermediates. The screens revealed SMARCA4 (SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4) as a stemness regulator, while balancing differentiation distinctively for each cell type. SMARCA4 knockdown in hematopoietic stem and progenitor cells caused impaired self-renewal in vitro and in vivo with skewed myeloid differentiation; whereas, in neural stem cells, it impaired self-renewal while biasing differentiation toward neural lineage, through combinatorial SWI/SNF subunit assembly. Our findings pose a powerful approach for deciphering human stem cell biology and attribute distinct roles to SMARCA4 in stem cell maintenance. : Güneş et al. show that RNAi screens on genetically identical (isogenic) human stem cells can dissect epigenetic factors important for self-renewal versus differentiation. By using iPSCs as a bridging cell type, the comparative RNAi screens in HSPCs and NSCs identify SMARCA4 as an important regulator of self-renewal and differentiation with cell-type-specific functions. Keywords: RNAi, comparative functional profiling, isogenic, hematopoiesis, self-renewal, neural differentiation, SMARCA4, SWI/SNFhttp://www.sciencedirect.com/science/article/pii/S2213671119300979 |
| spellingShingle | Ceren Güneş Maciej Paszkowski-Rogacz Susann Rahmig Shahryar Khattak Aylin Camgöz Martin Wermke Andreas Dahl Martin Bornhäuser Claudia Waskow Frank Buchholz Comparative RNAi Screens in Isogenic Human Stem Cells Reveal SMARCA4 as a Differential Regulator Stem Cell Reports |
| title | Comparative RNAi Screens in Isogenic Human Stem Cells Reveal SMARCA4 as a Differential Regulator |
| title_full | Comparative RNAi Screens in Isogenic Human Stem Cells Reveal SMARCA4 as a Differential Regulator |
| title_fullStr | Comparative RNAi Screens in Isogenic Human Stem Cells Reveal SMARCA4 as a Differential Regulator |
| title_full_unstemmed | Comparative RNAi Screens in Isogenic Human Stem Cells Reveal SMARCA4 as a Differential Regulator |
| title_short | Comparative RNAi Screens in Isogenic Human Stem Cells Reveal SMARCA4 as a Differential Regulator |
| title_sort | comparative rnai screens in isogenic human stem cells reveal smarca4 as a differential regulator |
| url | http://www.sciencedirect.com/science/article/pii/S2213671119300979 |
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