Identification and Characterization of the OCT4 Upstream Regulatory Region in Sus scrofa
OCT4 plays pivotal roles in maintaining pluripotency during early mammalian embryonic development and in embryonic stem cells. It is essential to establish a reporter system based on the OCT4 promoter region to study pluripotency. However, there is still a lack of information about the porcine OCT4...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2019/2130973 |
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| author | Seung-Hun Kim Kwang-Hwan Choi Dong-Kyung Lee Mingyun Lee Jae Yeon Hwang Chang-Kyu Lee |
| author_facet | Seung-Hun Kim Kwang-Hwan Choi Dong-Kyung Lee Mingyun Lee Jae Yeon Hwang Chang-Kyu Lee |
| author_sort | Seung-Hun Kim |
| collection | DOAJ |
| description | OCT4 plays pivotal roles in maintaining pluripotency during early mammalian embryonic development and in embryonic stem cells. It is essential to establish a reporter system based on the OCT4 promoter region to study pluripotency. However, there is still a lack of information about the porcine OCT4 upstream reporter system. To improve our understanding of the porcine OCT4 regulatory region, we identified conserved regions in the porcine OCT4 promoter upstream region by sequence-based comparative analysis using various mammalian genome sequences. The similarity of nucleotide sequences in the 5′ upstream region was low among mammalian species. However, the OCT4 promoter and four regulatory regions, including distal and proximal enhancer elements, had high similarity. Next, a functional analysis of the porcine OCT4 promoter region was conducted. Luciferase reporter assay results indicated that the porcine OCT4 distal enhancer and proximal enhancer were highly activated in mouse embryonic stem cells and embryonic carcinoma cells, respectively. A comparison analysis of naïve and primed state marker gene expression in a dual-reporter assay showed that the expression levels of naïve and primed markers differed in fluorescence signal between high-expressing cells and low-expressing cells. Similar to OCT4 upstream-based reporter systems derived from other species, the porcine OCT4 upstream region-based reporter constructs showed exclusive expression patterns depending on the state of pluripotency. This work provides basic information about the porcine OCT4 upstream region and various porcine OCT4 fluorescence reporter constructs, which can be applied to study species-specific pluripotency in early embryo development and the establishment of embryonic stem cells in pigs. |
| format | Article |
| id | doaj-art-d078b023f3d145ab8dea85f6603d8a19 |
| institution | OA Journals |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-d078b023f3d145ab8dea85f6603d8a192025-08-20T02:20:02ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/21309732130973Identification and Characterization of the OCT4 Upstream Regulatory Region in Sus scrofaSeung-Hun Kim0Kwang-Hwan Choi1Dong-Kyung Lee2Mingyun Lee3Jae Yeon Hwang4Chang-Kyu Lee5Department of Agricultural Biotechnology, Animal Biotechnology Major, Research Institute of Agriculture and Life Science, Seoul National University, Seoul 08826, Republic of KoreaDepartment of Agricultural Biotechnology, Animal Biotechnology Major, Research Institute of Agriculture and Life Science, Seoul National University, Seoul 08826, Republic of KoreaDepartment of Agricultural Biotechnology, Animal Biotechnology Major, Research Institute of Agriculture and Life Science, Seoul National University, Seoul 08826, Republic of KoreaDepartment of Agricultural Biotechnology, Animal Biotechnology Major, Research Institute of Agriculture and Life Science, Seoul National University, Seoul 08826, Republic of KoreaDepartment of Agricultural Biotechnology, Animal Biotechnology Major, Research Institute of Agriculture and Life Science, Seoul National University, Seoul 08826, Republic of KoreaDepartment of Agricultural Biotechnology, Animal Biotechnology Major, Research Institute of Agriculture and Life Science, Seoul National University, Seoul 08826, Republic of KoreaOCT4 plays pivotal roles in maintaining pluripotency during early mammalian embryonic development and in embryonic stem cells. It is essential to establish a reporter system based on the OCT4 promoter region to study pluripotency. However, there is still a lack of information about the porcine OCT4 upstream reporter system. To improve our understanding of the porcine OCT4 regulatory region, we identified conserved regions in the porcine OCT4 promoter upstream region by sequence-based comparative analysis using various mammalian genome sequences. The similarity of nucleotide sequences in the 5′ upstream region was low among mammalian species. However, the OCT4 promoter and four regulatory regions, including distal and proximal enhancer elements, had high similarity. Next, a functional analysis of the porcine OCT4 promoter region was conducted. Luciferase reporter assay results indicated that the porcine OCT4 distal enhancer and proximal enhancer were highly activated in mouse embryonic stem cells and embryonic carcinoma cells, respectively. A comparison analysis of naïve and primed state marker gene expression in a dual-reporter assay showed that the expression levels of naïve and primed markers differed in fluorescence signal between high-expressing cells and low-expressing cells. Similar to OCT4 upstream-based reporter systems derived from other species, the porcine OCT4 upstream region-based reporter constructs showed exclusive expression patterns depending on the state of pluripotency. This work provides basic information about the porcine OCT4 upstream region and various porcine OCT4 fluorescence reporter constructs, which can be applied to study species-specific pluripotency in early embryo development and the establishment of embryonic stem cells in pigs.http://dx.doi.org/10.1155/2019/2130973 |
| spellingShingle | Seung-Hun Kim Kwang-Hwan Choi Dong-Kyung Lee Mingyun Lee Jae Yeon Hwang Chang-Kyu Lee Identification and Characterization of the OCT4 Upstream Regulatory Region in Sus scrofa Stem Cells International |
| title | Identification and Characterization of the OCT4 Upstream Regulatory Region in Sus scrofa |
| title_full | Identification and Characterization of the OCT4 Upstream Regulatory Region in Sus scrofa |
| title_fullStr | Identification and Characterization of the OCT4 Upstream Regulatory Region in Sus scrofa |
| title_full_unstemmed | Identification and Characterization of the OCT4 Upstream Regulatory Region in Sus scrofa |
| title_short | Identification and Characterization of the OCT4 Upstream Regulatory Region in Sus scrofa |
| title_sort | identification and characterization of the oct4 upstream regulatory region in sus scrofa |
| url | http://dx.doi.org/10.1155/2019/2130973 |
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