A Stem Cell Reporter for Investigating Pluripotency and Self-Renewal in the Rat
Summary: Rat embryonic stem cells (rESCs) are capable of contributing to all differentiated tissues, including the germ line in chimeric animals, and represent a unique, authentic alternative to mouse embryonic stem cells for studying stem cell pluripotency and self-renewal. Here, we describe an EGF...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-01-01
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| Series: | Stem Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213671119304151 |
| _version_ | 1819076150927818752 |
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| author | Stephen Meek Jun Wei Taeho Oh Tom Watson Jaime Olavarrieta Linda Sutherland Daniel F. Carlson Angela Salzano Tamir Chandra Anagha Joshi Tom Burdon |
| author_facet | Stephen Meek Jun Wei Taeho Oh Tom Watson Jaime Olavarrieta Linda Sutherland Daniel F. Carlson Angela Salzano Tamir Chandra Anagha Joshi Tom Burdon |
| author_sort | Stephen Meek |
| collection | DOAJ |
| description | Summary: Rat embryonic stem cells (rESCs) are capable of contributing to all differentiated tissues, including the germ line in chimeric animals, and represent a unique, authentic alternative to mouse embryonic stem cells for studying stem cell pluripotency and self-renewal. Here, we describe an EGFP reporter transgene that tracks expression of the benchmark naive pluripotency marker gene Rex1 (Zfp42) in the rat. Insertion of the EGFP reporter gene downstream of the Rex1 promoter disrupted Rex1 expression, but REX1-deficient rESCs and rats were viable and apparently normal, validating this targeted knockin transgene as a neutral reporter. The Rex1-EGFP gene responded to self-renewal/differentiation factors and validated the critical role of β-catenin/LEF1 signaling. The stem cell reporter also allowed the identification of functionally distinct sub-populations of cells within rESC cultures, thus demonstrating its utility in discriminating between cell states in rat stem cell cultures, as well as providing a tool for tracking Rex1 expression in the rat. : Rat embryonic stem cells (ESCs) differentiate into all adult cells, including functional gametes, and uniquely qualify as a comparator with mouse ESCs to study self-renewal and pluripotency. In this article, Burdon and colleagues generated a Rex1-EGFP stem cell reporter as a sensitive tool to monitor and interrogate the regulation of rat ESCs, and contribute to understanding stem cells and fate determination in mammals. Keywords: rat, embryonic stem cell, Rex1, Zfp42, fluorescent reporter, EGFP, pluripotency, self-renewal, genetic engineering, transgenic, single cell sequencing |
| first_indexed | 2024-12-21T18:36:44Z |
| format | Article |
| id | doaj.art-73abfeba6940457291fa0e02eb90fc60 |
| institution | Directory Open Access Journal |
| issn | 2213-6711 |
| language | English |
| last_indexed | 2024-12-21T18:36:44Z |
| publishDate | 2020-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Reports |
| spelling | doaj.art-73abfeba6940457291fa0e02eb90fc602022-12-21T18:54:07ZengElsevierStem Cell Reports2213-67112020-01-01141154166A Stem Cell Reporter for Investigating Pluripotency and Self-Renewal in the RatStephen Meek0Jun Wei1Taeho Oh2Tom Watson3Jaime Olavarrieta4Linda Sutherland5Daniel F. Carlson6Angela Salzano7Tamir Chandra8Anagha Joshi9Tom Burdon10The Roslin Institute and R(D)VS, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UKThe Roslin Institute and R(D)VS, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UK; iRegene Therapeutics, C6-522, 666 Gaoxin Avenue, Wuhan, 430070, ChinaThe Roslin Institute and R(D)VS, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UKThe Roslin Institute and R(D)VS, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UKThe Roslin Institute and R(D)VS, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UKThe Roslin Institute and R(D)VS, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UKRecombinetics Inc., 1246 University Avenue W, St. Paul, MN 55125, USAMRC Unit for Human Genetics, Institute of Genetics and Molecular Medicine, The University of Edinburgh, Western General Hospital, Crewe Road South, Edinburgh, EH4 2XU, UKMRC Unit for Human Genetics, Institute of Genetics and Molecular Medicine, The University of Edinburgh, Western General Hospital, Crewe Road South, Edinburgh, EH4 2XU, UKThe Roslin Institute and R(D)VS, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UKThe Roslin Institute and R(D)VS, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UK; Corresponding authorSummary: Rat embryonic stem cells (rESCs) are capable of contributing to all differentiated tissues, including the germ line in chimeric animals, and represent a unique, authentic alternative to mouse embryonic stem cells for studying stem cell pluripotency and self-renewal. Here, we describe an EGFP reporter transgene that tracks expression of the benchmark naive pluripotency marker gene Rex1 (Zfp42) in the rat. Insertion of the EGFP reporter gene downstream of the Rex1 promoter disrupted Rex1 expression, but REX1-deficient rESCs and rats were viable and apparently normal, validating this targeted knockin transgene as a neutral reporter. The Rex1-EGFP gene responded to self-renewal/differentiation factors and validated the critical role of β-catenin/LEF1 signaling. The stem cell reporter also allowed the identification of functionally distinct sub-populations of cells within rESC cultures, thus demonstrating its utility in discriminating between cell states in rat stem cell cultures, as well as providing a tool for tracking Rex1 expression in the rat. : Rat embryonic stem cells (ESCs) differentiate into all adult cells, including functional gametes, and uniquely qualify as a comparator with mouse ESCs to study self-renewal and pluripotency. In this article, Burdon and colleagues generated a Rex1-EGFP stem cell reporter as a sensitive tool to monitor and interrogate the regulation of rat ESCs, and contribute to understanding stem cells and fate determination in mammals. Keywords: rat, embryonic stem cell, Rex1, Zfp42, fluorescent reporter, EGFP, pluripotency, self-renewal, genetic engineering, transgenic, single cell sequencinghttp://www.sciencedirect.com/science/article/pii/S2213671119304151 |
| spellingShingle | Stephen Meek Jun Wei Taeho Oh Tom Watson Jaime Olavarrieta Linda Sutherland Daniel F. Carlson Angela Salzano Tamir Chandra Anagha Joshi Tom Burdon A Stem Cell Reporter for Investigating Pluripotency and Self-Renewal in the Rat Stem Cell Reports |
| title | A Stem Cell Reporter for Investigating Pluripotency and Self-Renewal in the Rat |
| title_full | A Stem Cell Reporter for Investigating Pluripotency and Self-Renewal in the Rat |
| title_fullStr | A Stem Cell Reporter for Investigating Pluripotency and Self-Renewal in the Rat |
| title_full_unstemmed | A Stem Cell Reporter for Investigating Pluripotency and Self-Renewal in the Rat |
| title_short | A Stem Cell Reporter for Investigating Pluripotency and Self-Renewal in the Rat |
| title_sort | stem cell reporter for investigating pluripotency and self renewal in the rat |
| url | http://www.sciencedirect.com/science/article/pii/S2213671119304151 |
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