Inhibition of replication of goose paramyxovirus SF02 by hammerhead ribozyme targeting to the SF02 F mRNA in chicken embryo fibroblasts.
Hammerhead ribozyme RzF598 and its dysfunctional mutant dRzF598 targeting to the (F) gene of goose paramyxovirus SF02 have been designed. The transgenic plasmids pcDNA-RzF598 and pcDNA-dRzF598 were constructed by inserting ribozyme genes into eukaryotic expression vector pcDNA3. The plasmid pcDNA3 that lacks full ribozyme gene was used as a control. Plasmids pcDNA-RzF598, pcDNA-dRzF598 and pcDNA3 were transfected into chicken embryo fibroblasts (CEFs). The concentration of virus released by infected CEFs and the survival percentages of CEFs were identified. The results indicated that RzF598 successfully suppressed the replication of SF02 in CEFs. Survival percentage of CEFs being transfected with pcDNA-RzF598 and infected SF02 was up to 78.8%, while the survival percentages of untransfected CEFs and CEFs transfected with pcDNA3 after infection with SF02 were only about 5%.
Zou
Jian
J
Key laboratory of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, China.
Gong
Zun-Xun
ZX
D000818:Animals; D018109:Avulavirus / Q000235:genetics* / Q000254:growth & development; D002470:Cell Survival / Q000235:genetics; D002642:Chick Embryo; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D005777:Geese / Q000821:virology; D010957:Plasmids / Q000235:genetics; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014760:Viral Fusion Proteins / Q000235:genetics*; D014779:Virus Replication
D016337:RNA, Catalytic; D012333:RNA, Messenger; D012367:RNA, Viral; D014760:Viral Fusion Proteins; C427204:hammerhead ribozyme