Short hairpin RNA-mediated inhibition of measles virus replication in vitro.
Rab9 has been identified as a key component for the replication of measles virus (MV). In this study, gene-specific shRNAs were developed to suppress the replication of MV in culture cells by silencing the expression of Rab9 GTPase gene. Rab9 GTPase gene-specific shRNAs were designed and cloned into the expression vector of pSUPER.neo+EGFP. Vero-E6 cells were transfected with the recombinant plasmid via liposome and then infected with MV. The expression of Rab9 GTPase mRNA and protein were assayed by RT-PCR and Western blotting, respectively. ShRNA-mediated inhibition of MV replication was further evaluated by detecting the titer of MV. The results showed that the expression of Rab9 GTPase was dramatically and stably downregulated by the generated shRNAs targeting Rab9 GTPase gene, which contribute to the inhibition of MV replication, indicating these shRNAs could be potentially developed into therapeutic agents for the treatment of MV infection in the future.
Shi
Jun-Yan
JY
Department of Microbiology and Parasitology, College of Basic Medical Sciences, China Medical University, Shenyang 110001, PR China. junyanshi112@sina.com
Liu
Bing
B
Wang
Mei-Lian
ML
Luo
En-Jie
EJ
D000818:Animals; D002522:Chlorocebus aethiops; D015536:Down-Regulation; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D008457:Measles / Q000188:drug therapy / Q000821:virology; D008459:Measles virus / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D014709:Vero Cells; D014779:Virus Replication / Q000187:drug effects* / Q000235:genetics
D034741:RNA, Small Interfering