Tchurikov Nickolai A, Fedoseeva Daria M, Gashnikova Natalya M, Sosin Dmitri V, Gorbacheva Maria A, Alembekov Ildar R, Chechetkin Vladimir R, Kravatsky Yuri V, Kretova Olga V
Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
Engelhardt Institute of Molecular Biology, Moscow 119334, Russia.
Gene. 2016 May 25;583(1):78-83. doi: 10.1016/j.gene.2016.03.001. Epub 2016 Mar 3.
Highly active antiretroviral therapy has greatly reduced the morbidity and mortality of AIDS. However, many of the antiretroviral drugs are toxic with long-term use, and all currently used anti-HIV agents generate drug-resistant mutants. Therefore, there is a great need for new approaches to AIDS therapy. RNAi is a powerful means of inhibiting HIV-1 production in human cells. We propose to use RNAi for gene therapy of HIV/AIDS. Previously we identified a number of new biologically active siRNAs targeting several moderately conserved regions in HIV-1 transcripts. Here we analyze the heterogeneity of nucleotide sequences in three RNAi targets in sequences encoding the reverse transcriptase and integrase domains of current isolates of HIV-1 subtype A in Russia. These data were used to generate genetic constructs expressing short hairpin RNAs 28-30-bp in length that could be processed in cells into siRNAs. After transfection of the constructs we observed siRNAs that efficiently attacked the selected targets. We expect that targeting several viral genes important for HIV-1 reproduction will help overcome the problem of viral adaptation and will prevent the appearance of RNAi escape mutants in current virus strains, an important feature of gene therapy of HIV/AIDS.
高效抗逆转录病毒疗法已大大降低了艾滋病的发病率和死亡率。然而，许多抗逆转录病毒药物长期使用具有毒性，并且所有目前使用的抗HIV药物都会产生耐药突变体。因此，迫切需要新的艾滋病治疗方法。RNA干扰是抑制人类细胞中HIV-1产生的有力手段。我们提议将RNA干扰用于HIV/AIDS的基因治疗。此前我们鉴定了一些靶向HIV-1转录本中几个中度保守区域的新的具有生物活性的小干扰RNA。在此我们分析了俄罗斯目前HIV-1 A亚型分离株逆转录酶和整合酶结构域编码序列中三个RNA干扰靶点的核苷酸序列异质性。这些数据用于构建表达长度为28 - 30个碱基对的短发夹RNA的基因构建体，其可在细胞中加工成小干扰RNA。转染构建体后，我们观察到能有效攻击所选靶点的小干扰RNA。我们期望靶向几个对HIV-1复制重要的病毒基因将有助于克服病毒适应性问题，并防止在当前病毒株中出现RNA干扰逃逸突变体，这是HIV/AIDS基因治疗的一个重要特征。
