Role of circadian gene Clock during differentiation of mouse pluripotent stem cells
Abstract Biological rhythms controlled by the circadian clock are absent in embryonic stem cells (ESCs). However, they start to develop during the differentiation of pluripotent ESCs to downstream cells. Conversely, biological rhythms in adult somatic cells disappear when they are reprogrammed into...
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| Format: | Article |
| Language: | English |
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Oxford University Press
2016-09-01
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| Series: | Protein & Cell |
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| Online Access: | http://link.springer.com/article/10.1007/s13238-016-0319-9 |
| _version_ | 1797764386016722944 |
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| author | Chao Lu Yang Yang Ran Zhao Bingxuan Hua Chen Xu Zuoqin Yan Ning Sun Ruizhe Qian |
| author_facet | Chao Lu Yang Yang Ran Zhao Bingxuan Hua Chen Xu Zuoqin Yan Ning Sun Ruizhe Qian |
| author_sort | Chao Lu |
| collection | DOAJ |
| description | Abstract Biological rhythms controlled by the circadian clock are absent in embryonic stem cells (ESCs). However, they start to develop during the differentiation of pluripotent ESCs to downstream cells. Conversely, biological rhythms in adult somatic cells disappear when they are reprogrammed into induced pluripotent stem cells (iPSCs). These studies indicated that the development of biological rhythms in ESCs might be closely associated with the maintenance and differentiation of ESCs. The core circadian gene Clock is essential for regulation of biological rhythms. Its role in the development of biological rhythms of ESCs is totally unknown. Here, we used CRISPR/CAS9-mediated genetic editing techniques, to completely knock out the Clock expression in mouse ESCs. By AP, teratoma formation, quantitative real-time PCR and Immunofluorescent staining, we did not find any difference between Clock knockout mESCs and wild type mESCs in morphology and pluripotent capability under the pluripotent state. In brief, these data indicated Clock did not influence the maintaining of pluripotent state. However, they exhibited decreased proliferation and increased apoptosis. Furthermore, the biological rhythms failed to develop in Clock knockout mESCs after spontaneous differentiation, which indicated that there was no compensational factor in most peripheral tissues as described in mice models before (DeBruyne et al., 2007b). After spontaneous differentiation, loss of CLOCK protein due to Clock gene silencing induced spontaneous differentiation of mESCs, indicating an exit from the pluripotent state, or its differentiating ability. Our findings indicate that the core circadian gene Clock may be essential during normal mESCs differentiation by regulating mESCs proliferation, apoptosis and activity. |
| first_indexed | 2024-03-12T19:55:58Z |
| format | Article |
| id | doaj.art-752d7adfde974ab1b5896e5c1db7668c |
| institution | Directory Open Access Journal |
| issn | 1674-800X 1674-8018 |
| language | English |
| last_indexed | 2024-03-12T19:55:58Z |
| publishDate | 2016-09-01 |
| publisher | Oxford University Press |
| record_format | Article |
| series | Protein & Cell |
| spelling | doaj.art-752d7adfde974ab1b5896e5c1db7668c2023-08-02T02:49:52ZengOxford University PressProtein & Cell1674-800X1674-80182016-09-0171182083210.1007/s13238-016-0319-9Role of circadian gene Clock during differentiation of mouse pluripotent stem cellsChao Lu0Yang Yang1Ran Zhao2Bingxuan Hua3Chen Xu4Zuoqin Yan5Ning Sun6Ruizhe Qian7Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan UniversityDepartment of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan UniversityDepartment of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan UniversityDepartment of Orthopedics, Zhongshan Hospital, Fudan UniversityDepartment of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan UniversityDepartment of Orthopedics, Zhongshan Hospital, Fudan UniversityDepartment of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan UniversityDepartment of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan UniversityAbstract Biological rhythms controlled by the circadian clock are absent in embryonic stem cells (ESCs). However, they start to develop during the differentiation of pluripotent ESCs to downstream cells. Conversely, biological rhythms in adult somatic cells disappear when they are reprogrammed into induced pluripotent stem cells (iPSCs). These studies indicated that the development of biological rhythms in ESCs might be closely associated with the maintenance and differentiation of ESCs. The core circadian gene Clock is essential for regulation of biological rhythms. Its role in the development of biological rhythms of ESCs is totally unknown. Here, we used CRISPR/CAS9-mediated genetic editing techniques, to completely knock out the Clock expression in mouse ESCs. By AP, teratoma formation, quantitative real-time PCR and Immunofluorescent staining, we did not find any difference between Clock knockout mESCs and wild type mESCs in morphology and pluripotent capability under the pluripotent state. In brief, these data indicated Clock did not influence the maintaining of pluripotent state. However, they exhibited decreased proliferation and increased apoptosis. Furthermore, the biological rhythms failed to develop in Clock knockout mESCs after spontaneous differentiation, which indicated that there was no compensational factor in most peripheral tissues as described in mice models before (DeBruyne et al., 2007b). After spontaneous differentiation, loss of CLOCK protein due to Clock gene silencing induced spontaneous differentiation of mESCs, indicating an exit from the pluripotent state, or its differentiating ability. Our findings indicate that the core circadian gene Clock may be essential during normal mESCs differentiation by regulating mESCs proliferation, apoptosis and activity.http://link.springer.com/article/10.1007/s13238-016-0319-9Circadian gene Clockmouse embryonic stem cellsgene knockoutpluripotencycell proliferationcell apoptosis |
| spellingShingle | Chao Lu Yang Yang Ran Zhao Bingxuan Hua Chen Xu Zuoqin Yan Ning Sun Ruizhe Qian Role of circadian gene Clock during differentiation of mouse pluripotent stem cells Protein & Cell Circadian gene Clock mouse embryonic stem cells gene knockout pluripotency cell proliferation cell apoptosis |
| title | Role of circadian gene Clock during differentiation of mouse pluripotent stem cells |
| title_full | Role of circadian gene Clock during differentiation of mouse pluripotent stem cells |
| title_fullStr | Role of circadian gene Clock during differentiation of mouse pluripotent stem cells |
| title_full_unstemmed | Role of circadian gene Clock during differentiation of mouse pluripotent stem cells |
| title_short | Role of circadian gene Clock during differentiation of mouse pluripotent stem cells |
| title_sort | role of circadian gene clock during differentiation of mouse pluripotent stem cells |
| topic | Circadian gene Clock mouse embryonic stem cells gene knockout pluripotency cell proliferation cell apoptosis |
| url | http://link.springer.com/article/10.1007/s13238-016-0319-9 |
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