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Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish
Su, Jianguo1,2; Zhu, Zuoyan1; Xiong, Feng1; Wang, Yaping1; Zhu, ZY, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2008-09-01
Source PublicationMARINE BIOTECHNOLOGY
ISSN1436-2228
Volume10Issue:5Pages:511-517
AbstractShort 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.; 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.
SubtypeArticle
KeywordHybrid Promoter Cmv Promoter U6 Promoter Rnai Zebrafish
Department[Su, Jianguo; Zhu, Zuoyan; Xiong, Feng; Wang, Yaping] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; [Su, Jianguo] NW A&F Univ, Coll Anim Sci & Technol, Dept Aquaculture, Yangling 712100, Peoples R China
Subject AreaBiotechnology & Applied Microbiology ; Marine & Freshwater Biology
DOI10.1007/s10126-008-9087-8
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaBiotechnology & Applied Microbiology ; Marine & Freshwater Biology
WOS SubjectBiotechnology & Applied Microbiology ; Marine & Freshwater Biology
WOS IDWOS:000258323900005
WOS KeywordDOUBLE-STRANDED-RNA ; POLYMERASE-III ; MAMMALIAN-CELLS ; BINDING-PROTEIN ; GENE ; TRANSCRIPTION ; EXPRESSION ; INJECTION ; EMBRYOS ; SNRNA
Citation statistics
Cited Times:10[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8038
Collection期刊论文
Corresponding AuthorZhu, ZY, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Affiliation1.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
Recommended Citation
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,&Zhu, ZY, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(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.
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