Inhibition of retroviral pathogenesis by RNA interference
Inhibition of retroviral pathogenesis by RNA interference
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DOI：
10.1016/s0960-9822(02)00975-2
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发表时间：
2002-08-06
期刊：
影响因子：
9.2
通讯作者：
Bushman, FD
中科院分区：
文献类型：
--
作者：
Hu, WY;Myers, CP;Bushman, FD
背景资料：RNA干扰（RNAi）是一种新发现的细胞防御系统，已知其在模式生物中抑制基因组寄生虫的复制。结果：Rous肉瘤病毒（Rous sarcoma virus，RSV）前病毒DNA经电穿孔进入发育中的鸡神经管后，可启动逆转录病毒感染。共电穿孔的前病毒DNA和短的双链RNA匹配序列的禽流感逆转录病毒，这是设计来诱导RNAi抗RSV，抑制病毒复制。电穿孔后RSV的复制导致胚胎发育中断和早期死亡，但这也可以通过针对RSV基因组的RNAi来抑制。RNAi还可以抑制RSV和HIV在细胞培养中的生长。对被RNAi抑制的逆转录病毒生命周期步骤的分析表明，它主要阻止了感染后期合成的病毒RNA的积累。结论：RNAi可以阻断脊椎动物的逆转录病毒感染。本文所述的组织电穿孔方法将使RNAi广泛用于研究脊椎动物的基因功能和感染控制。
Background: RNA interference (RNAi) is a newly discovered cellular defense system that is known to suppress replication of genomic parasites in model organisms. It has been widely conjectured that RNAi may also serve as an antiviral system in vertebrates.Results: Retroviral infection could be initiated by electroporation of cloned Rous sarcoma virus (RSV) proviral DNA into the developing chick neural tube. Coelectroporation of proviral DNA and short double-stranded RNAs matching sequences of avain retroviruses, which were designed to induce RNAi against RSV, inhibited viral replication. Replication of RSV after electroporation resulted in disruption of embryonic development and early death, but this, too, could be suppressed by RNAi against the RSV genome. RNAi could also inhibit the growth of RSV and HIV in cell culture. Analysis of the step of the retroviral life cycle that is inhibited by RNAi revealed that it primarily prevented accumulation of the viral RNAs synthesized late during infection. RNA genomes introduced in viral particles early during infection were less sensitive.Conclusions: RNAi can block retroviral infection in vertebrates. The tissue electroporation method described here should allow RNAi to be used widely to study gene function and control of infection in vertebrate animals.