Department of Microbiology and Parasitology, College of Basic Medical Sciences, China Medical University, Shenyang 110001, PR China.
Can J Microbiol. 2010 Jan;56(1):77-80. doi: 10.1139/w09-103.
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.
Rab9 已被确定为麻疹病毒 (MV) 复制的关键组成部分。在这项研究中,通过沉默 Rab9 GTPase 基因的表达,开发了基因特异性 shRNA,以抑制 MV 在培养细胞中的复制。设计了 Rab9 GTPase 基因特异性 shRNA,并将其克隆到 pSUPER.neo+EGFP 表达载体中。通过脂质体转染 Vero-E6 细胞,并随后用 MV 感染。通过 RT-PCR 和 Western blot 分别检测 Rab9 GTPase mRNA 和蛋白质的表达。通过检测 MV 的滴度进一步评估 shRNA 介导的 MV 复制抑制。结果表明,针对 Rab9 GTPase 基因生成的 shRNA 可显著且稳定地下调 Rab9 GTPase 的表达,这有助于抑制 MV 复制,表明这些 shRNA 将来可能被开发为治疗 MV 感染的治疗剂。