Lentiviral-mediated delivery of combined HIV-1 decoy TAR and Vif siRNA as a single RNA molecule that cleaves to inhibit HIV-1 in transduced cells
Lentiviral-mediated delivery of combined HIV-1 decoy TAR and Vif siRNA as a single RNA molecule that cleaves to inhibit HIV-1 in transduced cells
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DOI：
10.1081/ncn-200059981
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发表时间：
2005-01-01
影响因子：
1.3
通讯作者：
Takaku, H
中科院分区：
文献类型：
--
作者：
Barnor, JS;Miyano-Kurosaki, N;Takaku, H
RNA干扰（RNA（i））通过短的干扰21 - 23 mer双链RNA（siRNA）片段沉默基因表达，所述siRNA片段以序列特异性方式引导同源mRNA降解。另一方面，已知HIV-1诱饵TAR RNA与HIV-1达特蛋白竞争性相互作用，以下调来自长末端重复序列（LTR）启动子的增强的基因表达。在这里，我们报告了一种新的表达构建体，编码HIV-1诱饵TAR和Vif siRNA，作为单一的RNA底物，在人U6启动子的控制下表达，然后TAR和siRNA被内源性RNase HT样酶切割成各自独立的RNA。然后，每个切割的HIV-1抗基因协同地有助于增强转导的Jurkat细胞中HIV-1复制的抑制功效（> 80%）。这些结果表明，用同时表达的细胞内诱饵TAR和Vif-siRNA靶向HIV-I mRNA可以导致用于控制和管理HIV-AIDS的有效基因治疗策略。
RNA interference (RNA(i)) silences gene expression via short interfering 21-23 mer double-stranded RNA (siRNA) segments that guide cognate mRNA degradation in a sequence-specific manner. On the other hand, HIV-1 decoy TAR RNA are known to competitively interact with the HIV-1 Tat protein, to downregulate the enhanced gene expression from the long terminal repeat (LTR) promoters. Here we report that a novel expression construct, encoding both HIV-1 decoy TAR and Vif siRNA, as a single RNA substrate, was expressed under the control of the human U6 promoter, and later the TAR and siRNA were cleaved into their respective separate RNA by the endogenous RNase HT like enzyme. Each of the cleaved HIV-1 anti-genes then synergistically contributed toward enhancing the inhibition efficacy (>80%) of HIV-1 replication in transduced Jurkat cells. These results suggest that targeting HIV- I mRNA with simultaneously expressed intracellular decoy TAR and Vif-siRNA could lead to an effective gene therapy strategy for the control and management of HIV-AIDS.