Boden Daniel, Pusch Oliver, Silbermann Rebecca, Lee Fred, Tucker Lynne, Ramratnam Bharat
Laboratory of Retrovirology, Division of Infectious Diseases, Department of Medicine, Brown Medical School, 4th Floor, 55 Claverick Street, Providence, RI 02903, USA.
Nucleic Acids Res. 2004 Feb 13;32(3):1154-8. doi: 10.1093/nar/gkh278. Print 2004.
Post-transcriptional inhibition of HIV-1 replication can be achieved by RNA interference (RNAi). The cellular expression of short interfering RNA (siRNA) or short hairpin RNA (shRNA) homologous to regions of the HIV-1 genome decreases viral replication by the selective degradation of targeted RNA. Here, we demonstrate that another class of noncoding regulatory RNA, termed microRNA (miRNA), can be used to deliver antiviral RNAi. By incorporating sequences encoding siRNA targeting the HIV-1 transactivator protein tat into a human miR-30 pre-microRNA (pre-miRNA) backbone, we were able to express tat siRNA in cells. The tat siRNA delivered as pre-miRNA precursor was 80% more effective in reducing HIV-1 p24 antigen production than tat siRNA expressed as conventional shRNA. Our results confirm the utility of expressing HIV-1 specific siRNA through a miR-30 precursor stem-loop structure and suggest that this strategy can be used to increase the antiviral potency of RNAi.
RNA干扰（RNAi）可实现对HIV-1复制的转录后抑制。与HIV-1基因组区域同源的短干扰RNA（siRNA）或短发夹RNA（shRNA）的细胞表达通过靶向RNA的选择性降解降低病毒复制。在此，我们证明另一类非编码调节RNA，即微RNA（miRNA），可用于传递抗病毒RNAi。通过将编码靶向HIV-1反式激活蛋白tat的siRNA的序列整合到人miR-30前体微RNA（pre-miRNA）骨架中，我们能够在细胞中表达tat siRNA。作为pre-miRNA前体传递的tat siRNA在降低HIV-1 p24抗原产生方面比作为传统shRNA表达的tat siRNA有效80%。我们的结果证实了通过miR-30前体茎环结构表达HIV-1特异性siRNA的实用性，并表明该策略可用于提高RNAi的抗病毒效力。
