Avian influenza virus H5N1 causes widespread infection in the birds and human respiratory tract, but existing vaccines and drug therapy are of limited value. Here we show that small interfering RNAs (siRNAs) specific for conserved regions of the viral genome can potently inhibit influenza virus production in cell lines, embryonated chicken eggs and BALB/c mice. siRNA expression plasmid pBabe-Super was chosen in the study, which directed the synthesis of small interfering RNAs in cells. The inhibition depended on the presence of a functional antisense strand in the small interfering RNA duplex, suggesting that viral mRNA is the target of RNA interference (RNAi). Among the three small interfering RNA expression plasmids we designed, we found that small interfering RNA for nucleocapsid protein (NP) had a specific effect in inhibiting the accumulation of RNAs in infected cells because of a critical requirement for newly synthesized nucleocapsid proteins in avian influenza viral RNA transcription and replication. The findings reveal that newly synthesized nucleocapsid, polymerase A (PA) and polymerase B1 (PB1) proteins are required for avian influenza virus transcription and replication and provide a basis for the development of small interfering RNAs as prophylaxis and therapy for avian influenza infection in birds and humans.

译文

:H5N1禽流感病毒在鸟类和人类呼吸道中引起广泛感染,但现有的疫苗和药物疗法的价值有限。在这里,我们显示了特异性针对病毒基因组保守区的小干扰RNA(siRNA)可以有效抑制细胞系,胚胎鸡卵和BALB / c小鼠中的流感病毒产生。在研究中选择了siRNA表达质粒pBabe-Super,该质粒指导细胞中小的干扰RNA的合成。抑制取决于小干扰RNA双链体中功能性反义链的存在,表明病毒mRNA是RNA干扰(RNAi)的靶标。在我们设计的三种小分子干扰RNA表达质粒中,我们发现对于核衣壳蛋白(NP)的小分子干扰RNA在抑制感染细胞中RNA的积累方面具有特定作用,因为对禽流感病毒中新合成的核衣壳蛋白的关键需求RNA转录和复制。研究结果表明,新合成的核衣壳,聚合酶A(PA)和聚合酶B1(PB1)蛋白是禽流感病毒转录和复制所必需的,并为开发小干扰RNA作为预防和治疗禽流感的方法奠定了基础。和人类。

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