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Cped1 promotes chicken SSCs formation with the aid of histone acetylation and transcription factor Sox2
Spermatogonial stem cells (SSCs) may apply to gene therapy, regenerative medicine in place of embryonic stem cells (ESCs). However, the application of SSCs was severely limited by the low induction efficiency and the lack of thorough analysis of the regulatory mechanisms of SSCs formation. Current e...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6137251/ https://www.ncbi.nlm.nih.gov/pubmed/30038055 http://dx.doi.org/10.1042/BSR20180707 |
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author | Zhang, Chen Wang, Fei Zuo, Qisheng Sun, Changhua Jin, Jing Li, Tingting Wang, Man Zhao, Ruifeng Yu, Xinjian Sun, Hongyan Zhang, Yani Chen, GuoHong Li, Bichun |
author_facet | Zhang, Chen Wang, Fei Zuo, Qisheng Sun, Changhua Jin, Jing Li, Tingting Wang, Man Zhao, Ruifeng Yu, Xinjian Sun, Hongyan Zhang, Yani Chen, GuoHong Li, Bichun |
author_sort | Zhang, Chen |
collection | PubMed |
description | Spermatogonial stem cells (SSCs) may apply to gene therapy, regenerative medicine in place of embryonic stem cells (ESCs). However, the application of SSCs was severely limited by the low induction efficiency and the lack of thorough analysis of the regulatory mechanisms of SSCs formation. Current evidences have demonstrated multiple marker genes of germ cells, while genes that specifically regulate the formation of SSCs have not been explored. In our study, cadherin-like and PC-esterase domain containing 1 (Cped1) expressed specifically in SSCs based on RNA-seq data analysis. To study the function of Cped1 in the formation of SSCs, we successfully established a CRISPR/Cas9 knockout system. The gene disruption frequency is 37% in DF1 and 25% in ESCs without off-target effects. Knockout of Cped1 could significantly inhibit the formation of SSCs in vivo and in vitro. The fragment of −1050 to −1 bp had the activity as Cped1 gene promoter. Histone acetylation could regulate the expression of Cped1. We added 5-azaeytidi (DNA methylation inhibitors) and TSA (histone deacetylase inhibitors) respectively during the cultivation of SSCs. TSA was validated to promote the transcription of Cped1. Dual-luciferase reporter assay revealed that active control area of the chicken Cped1 gene is −296 to −1 bp. There are Cebpb, Sp1, and Sox2 transcription factor binding sites in this region. Point-mutation experiment results showed that Sox2 negatively regulates the transcription of Cped1. Above results demonstrated that Cped1 is a key gene that regulates the formation of SSCs. Histone acetylation and transcription factor Sox2 participate in the regulation of Cped1. |
format | Online Article Text |
id | pubmed-6137251 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-61372512018-09-18 Cped1 promotes chicken SSCs formation with the aid of histone acetylation and transcription factor Sox2 Zhang, Chen Wang, Fei Zuo, Qisheng Sun, Changhua Jin, Jing Li, Tingting Wang, Man Zhao, Ruifeng Yu, Xinjian Sun, Hongyan Zhang, Yani Chen, GuoHong Li, Bichun Biosci Rep Research Articles Spermatogonial stem cells (SSCs) may apply to gene therapy, regenerative medicine in place of embryonic stem cells (ESCs). However, the application of SSCs was severely limited by the low induction efficiency and the lack of thorough analysis of the regulatory mechanisms of SSCs formation. Current evidences have demonstrated multiple marker genes of germ cells, while genes that specifically regulate the formation of SSCs have not been explored. In our study, cadherin-like and PC-esterase domain containing 1 (Cped1) expressed specifically in SSCs based on RNA-seq data analysis. To study the function of Cped1 in the formation of SSCs, we successfully established a CRISPR/Cas9 knockout system. The gene disruption frequency is 37% in DF1 and 25% in ESCs without off-target effects. Knockout of Cped1 could significantly inhibit the formation of SSCs in vivo and in vitro. The fragment of −1050 to −1 bp had the activity as Cped1 gene promoter. Histone acetylation could regulate the expression of Cped1. We added 5-azaeytidi (DNA methylation inhibitors) and TSA (histone deacetylase inhibitors) respectively during the cultivation of SSCs. TSA was validated to promote the transcription of Cped1. Dual-luciferase reporter assay revealed that active control area of the chicken Cped1 gene is −296 to −1 bp. There are Cebpb, Sp1, and Sox2 transcription factor binding sites in this region. Point-mutation experiment results showed that Sox2 negatively regulates the transcription of Cped1. Above results demonstrated that Cped1 is a key gene that regulates the formation of SSCs. Histone acetylation and transcription factor Sox2 participate in the regulation of Cped1. Portland Press Ltd. 2018-09-14 /pmc/articles/PMC6137251/ /pubmed/30038055 http://dx.doi.org/10.1042/BSR20180707 Text en © 2018 The Author(s). http://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (http://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Articles Zhang, Chen Wang, Fei Zuo, Qisheng Sun, Changhua Jin, Jing Li, Tingting Wang, Man Zhao, Ruifeng Yu, Xinjian Sun, Hongyan Zhang, Yani Chen, GuoHong Li, Bichun Cped1 promotes chicken SSCs formation with the aid of histone acetylation and transcription factor Sox2 |
title | Cped1 promotes chicken SSCs formation with the aid of histone acetylation and transcription factor Sox2 |
title_full | Cped1 promotes chicken SSCs formation with the aid of histone acetylation and transcription factor Sox2 |
title_fullStr | Cped1 promotes chicken SSCs formation with the aid of histone acetylation and transcription factor Sox2 |
title_full_unstemmed | Cped1 promotes chicken SSCs formation with the aid of histone acetylation and transcription factor Sox2 |
title_short | Cped1 promotes chicken SSCs formation with the aid of histone acetylation and transcription factor Sox2 |
title_sort | cped1 promotes chicken sscs formation with the aid of histone acetylation and transcription factor sox2 |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6137251/ https://www.ncbi.nlm.nih.gov/pubmed/30038055 http://dx.doi.org/10.1042/BSR20180707 |
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