Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish
文献类型：期刊论文
| 作者 | Su, Jianguo1,2; Zhu, Zuoyan1; Xiong, Feng1; Wang, Yaping1 |
| 刊名 | MARINE BIOTECHNOLOGY
 |
| 出版日期 | 2008-09-01 |
| 卷号 | 10期号:5页码:511-517 |
| ISSN号 | 1436-2228 |
| 关键词 | hybrid promoter CMV promoter U6 promoter RNAi zebrafish |
| 通讯作者 | Zhu, ZY, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China |
| 中文摘要 | Short hairpin RNA (shRNA) directed by RNA polymerase III (Pol III) or Pol II promoter was shown to be capable of silencing gene expression, which should permit analyses of gene functions or as a potential therapeutic tool. However, the inhibitory effect of shRNA remains problematic in fish. We demonstrated that silencing efficiency by shRNA produced from the hybrid construct composed of the CMV enhancer or entire CMV promoter placed immediately upstream of a U6 promoter. When tested the exogenous gene, silencing of an enhanced green fluorescent protein (EGFP) target gene was 89.18 +/- 5.06% for CMVE-U6 promoter group and 88.26 +/- 6.46% for CMV-U6 promoter group. To test the hybrid promoters driving shRNA efficiency against an endogenous gene, we used shRNA against no tail (NTL) gene. When vectorized in the zebrafish, the hybrid constructs strongly repressed NTL gene expression. The NTL phenotype occupied 52.09 +/- 3.06% and 51.56 +/- 3.68% for CMVE-U6 promoter and CMV-U6 promoter groups, respectively. The NTL gene expression reduced 82.17 +/- 2.96% for CMVE-U6 promoter group and 83.06 +/- 2.38% for CMV-U6 promoter group. We concluded that the CMV enhancer or entire CMV promoter locating upstream of the U6-promoter could significantly improve inhibitory effect induced by the shRNA for both exogenous and endogenous genes compared with the CMV promoter or U6 promoter alone. In contrast, the two hybrid promoter constructs had similar effects on driving shRNA. |
| 英文摘要 | Short hairpin RNA (shRNA) directed by RNA polymerase III (Pol III) or Pol II promoter was shown to be capable of silencing gene expression, which should permit analyses of gene functions or as a potential therapeutic tool. However, the inhibitory effect of shRNA remains problematic in fish. We demonstrated that silencing efficiency by shRNA produced from the hybrid construct composed of the CMV enhancer or entire CMV promoter placed immediately upstream of a U6 promoter. When tested the exogenous gene, silencing of an enhanced green fluorescent protein (EGFP) target gene was 89.18 +/- 5.06% for CMVE-U6 promoter group and 88.26 +/- 6.46% for CMV-U6 promoter group. To test the hybrid promoters driving shRNA efficiency against an endogenous gene, we used shRNA against no tail (NTL) gene. When vectorized in the zebrafish, the hybrid constructs strongly repressed NTL gene expression. The NTL phenotype occupied 52.09 +/- 3.06% and 51.56 +/- 3.68% for CMVE-U6 promoter and CMV-U6 promoter groups, respectively. The NTL gene expression reduced 82.17 +/- 2.96% for CMVE-U6 promoter group and 83.06 +/- 2.38% for CMV-U6 promoter group. We concluded that the CMV enhancer or entire CMV promoter locating upstream of the U6-promoter could significantly improve inhibitory effect induced by the shRNA for both exogenous and endogenous genes compared with the CMV promoter or U6 promoter alone. In contrast, the two hybrid promoter constructs had similar effects on driving shRNA. |
| 学科主题 | Biotechnology & Applied Microbiology; Marine & Freshwater Biology |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology ; Marine & Freshwater Biology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology ; Marine & Freshwater Biology |
| 关键词[WOS] | DOUBLE-STRANDED-RNA ; POLYMERASE-III ; MAMMALIAN-CELLS ; BINDING-PROTEIN ; GENE ; TRANSCRIPTION ; EXPRESSION ; INJECTION ; EMBRYOS ; SNRNA |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000258323900005 |
| 公开日期 | 2010-10-13 |
| 源URL | [http://ir.ihb.ac.cn/handle/152342/8038]  |
| 专题 | 水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文 |
| 作者单位 | 1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China 2.NW A&F Univ, Coll Anim Sci & Technol, Dept Aquaculture, Yangling 712100, Peoples R China |
| 推荐引用方式 GB/T 7714 | Su, Jianguo,Zhu, Zuoyan,Xiong, Feng,et al. Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish[J]. MARINE BIOTECHNOLOGY,2008,10(5):511-517. |
| APA | Su, Jianguo,Zhu, Zuoyan,Xiong, Feng,&Wang, Yaping.(2008).Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish.MARINE BIOTECHNOLOGY,10(5),511-517. |
| MLA | Su, Jianguo,et al."Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish".MARINE BIOTECHNOLOGY 10.5(2008):511-517. |
入库方式： OAI收割
来源：水生生物研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。