Efficient Gene Transfer and Gene Editing in Sterlet (Acipenser ruthenus)
The sturgeon (Acipenseriformes) is an important farmed species because of its economical value. However, neither gene transfer nor gene editing techniques have been established in sturgeon for molecular breeding and gene functional study until now. In this study, we accomplished gene transfer and ge...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2018-04-01
|
| Series: | Frontiers in Genetics |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/article/10.3389/fgene.2018.00117/full |
| _version_ | 1818507145822339072 |
|---|---|
| author | Ji Chen Wei Wang Zhaohui Tian Ying Dong Tian Dong Hua Zhu Zuoyan Zhu Hongxia Hu Wei Hu Wei Hu |
| author_facet | Ji Chen Wei Wang Zhaohui Tian Ying Dong Tian Dong Hua Zhu Zuoyan Zhu Hongxia Hu Wei Hu Wei Hu |
| author_sort | Ji Chen |
| collection | DOAJ |
| description | The sturgeon (Acipenseriformes) is an important farmed species because of its economical value. However, neither gene transfer nor gene editing techniques have been established in sturgeon for molecular breeding and gene functional study until now. In this study, we accomplished gene transfer and gene editing in sterlet (Acipenser ruthenus), which has the shortest sexual maturation period of sturgeons. The plasmid encoding enhanced green fluorescent protein (EGFP) was transferred into the embryos of sterlet at injection concentration of 100 ng/μL, under which condition high survival rate and gene transfer rate could be achieved. Subsequently, exogenous EGFP was efficiently disrupted by transcription activator-like effector nucleases (TALENs) or clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 nuclease/guide RNA (gRNA), with injection concentrations of 300 ng/μL TALENs, or 100 ng/μL Cas9 nuclease and 30 ng/μL gRNA, respectively, under which condition high survival rate and gene mutation rate could be achieved. Finally, the endogenous gene no tail in sterlet was successfully mutated by Cas9 nuclease/gRNA. We observed the CRISPR-induced no tail mutation, at a high efficiency with the mutant P0 embryos displaying the expected phenotype of bent spine and twisted tail. |
| first_indexed | 2024-12-10T22:14:34Z |
| format | Article |
| id | doaj.art-354071a60a8e445e9c4fa2f1581d3d9c |
| institution | Directory Open Access Journal |
| issn | 1664-8021 |
| language | English |
| last_indexed | 2024-12-10T22:14:34Z |
| publishDate | 2018-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Genetics |
| spelling | doaj.art-354071a60a8e445e9c4fa2f1581d3d9c2022-12-22T01:31:30ZengFrontiers Media S.A.Frontiers in Genetics1664-80212018-04-01910.3389/fgene.2018.00117345219Efficient Gene Transfer and Gene Editing in Sterlet (Acipenser ruthenus)Ji Chen0Wei Wang1Zhaohui Tian2Ying Dong3Tian Dong4Hua Zhu5Zuoyan Zhu6Hongxia Hu7Wei Hu8Wei Hu9State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaBeijing Fisheries Research Institute, Beijing Key Laboratory of Fishery Biotechnology, Beijing, ChinaBeijing Fisheries Research Institute, Beijing Key Laboratory of Fishery Biotechnology, Beijing, ChinaBeijing Fisheries Research Institute, Beijing Key Laboratory of Fishery Biotechnology, Beijing, ChinaBeijing Fisheries Research Institute, Beijing Key Laboratory of Fishery Biotechnology, Beijing, ChinaBeijing Fisheries Research Institute, Beijing Key Laboratory of Fishery Biotechnology, Beijing, ChinaState Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaBeijing Fisheries Research Institute, Beijing Key Laboratory of Fishery Biotechnology, Beijing, ChinaState Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, ChinaQingdao National Laboratory for Marine Science and Technology, Qingdao, ChinaThe sturgeon (Acipenseriformes) is an important farmed species because of its economical value. However, neither gene transfer nor gene editing techniques have been established in sturgeon for molecular breeding and gene functional study until now. In this study, we accomplished gene transfer and gene editing in sterlet (Acipenser ruthenus), which has the shortest sexual maturation period of sturgeons. The plasmid encoding enhanced green fluorescent protein (EGFP) was transferred into the embryos of sterlet at injection concentration of 100 ng/μL, under which condition high survival rate and gene transfer rate could be achieved. Subsequently, exogenous EGFP was efficiently disrupted by transcription activator-like effector nucleases (TALENs) or clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 nuclease/guide RNA (gRNA), with injection concentrations of 300 ng/μL TALENs, or 100 ng/μL Cas9 nuclease and 30 ng/μL gRNA, respectively, under which condition high survival rate and gene mutation rate could be achieved. Finally, the endogenous gene no tail in sterlet was successfully mutated by Cas9 nuclease/gRNA. We observed the CRISPR-induced no tail mutation, at a high efficiency with the mutant P0 embryos displaying the expected phenotype of bent spine and twisted tail.http://journal.frontiersin.org/article/10.3389/fgene.2018.00117/fullsterletgene transfergene editingEGFPno tail |
| spellingShingle | Ji Chen Wei Wang Zhaohui Tian Ying Dong Tian Dong Hua Zhu Zuoyan Zhu Hongxia Hu Wei Hu Wei Hu Efficient Gene Transfer and Gene Editing in Sterlet (Acipenser ruthenus) Frontiers in Genetics sterlet gene transfer gene editing EGFP no tail |
| title | Efficient Gene Transfer and Gene Editing in Sterlet (Acipenser ruthenus) |
| title_full | Efficient Gene Transfer and Gene Editing in Sterlet (Acipenser ruthenus) |
| title_fullStr | Efficient Gene Transfer and Gene Editing in Sterlet (Acipenser ruthenus) |
| title_full_unstemmed | Efficient Gene Transfer and Gene Editing in Sterlet (Acipenser ruthenus) |
| title_short | Efficient Gene Transfer and Gene Editing in Sterlet (Acipenser ruthenus) |
| title_sort | efficient gene transfer and gene editing in sterlet acipenser ruthenus |
| topic | sterlet gene transfer gene editing EGFP no tail |
| url | http://journal.frontiersin.org/article/10.3389/fgene.2018.00117/full |
| work_keys_str_mv | AT jichen efficientgenetransferandgeneeditinginsterletacipenserruthenus AT weiwang efficientgenetransferandgeneeditinginsterletacipenserruthenus AT zhaohuitian efficientgenetransferandgeneeditinginsterletacipenserruthenus AT yingdong efficientgenetransferandgeneeditinginsterletacipenserruthenus AT tiandong efficientgenetransferandgeneeditinginsterletacipenserruthenus AT huazhu efficientgenetransferandgeneeditinginsterletacipenserruthenus AT zuoyanzhu efficientgenetransferandgeneeditinginsterletacipenserruthenus AT hongxiahu efficientgenetransferandgeneeditinginsterletacipenserruthenus AT weihu efficientgenetransferandgeneeditinginsterletacipenserruthenus AT weihu efficientgenetransferandgeneeditinginsterletacipenserruthenus |