Abstract
Rat embryonic stem cells (rESCs) are capable of contributing to all differentiated tissues including the germ line in chimaeric animals, and represent a unique, authentic, alternative to mouse ESCs for studying stem cell pluripotency and self-renewal. Here we describe an EGFP reporter transgene that tracks expression of the benchmark naïve 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 knock-in transgene as a neutral reporter. The Rex1-EGFP gene responded to self-renewal/differentiation factors and validated the critical role of β- catenin/LEF1 signalling. 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.
disrupted Rex1 expression, but REX1-deficient rESCs and rats were viable and apparently normal, validating this targeted knock-in transgene as a neutral reporter. The Rex1-EGFP gene responded to self-renewal/differentiation factors and validated the critical role of β- catenin/LEF1 signalling. 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.
Original language | English |
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Journal | Stem Cell Reports |
Early online date | 2 Jan 2020 |
DOIs | |
Publication status | E-pub ahead of print - 2 Jan 2020 |
Keywords / Materials (for Non-textual outputs)
- rat
- embryonic stem cell
- Rex1
- Zfp42
- fluorescent reporter
- pluripotency
- self-renewal
- genetic engineering
- transgenic
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Single cell transcriptional profiling of rat embryonic stem cells defined by Rex1-EGFP reporter expression
Meek, S. (Creator), Chandra, T. (Creator), Joshi, A. (Creator) & Burdon, T. (Creator), Edinburgh DataShare, 1 Nov 2019
DOI: 10.7488/ds/2639
Dataset
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Primary Dataset of Single cell transcriptional profiling of rat embryonic stem cells defined by Rex1-EGFP reporter expression
Burdon, T. (Creator), Meek, S. (Creator), Chandra, T. (Creator), Joshi, A. (Creator) & Simpson, C. (Data Manager), Edinburgh DataVault, 2020
DOI: 10.7488/078deee1-6e1e-4bae-a483-4ec093ec126f
Dataset
Profiles
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Tom Burdon
- Royal (Dick) School of Veterinary Studies - Group Leader/Senior Research Fellow
Person: Academic: Research Active