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.
RNA interference (RNAi) 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 III-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-1 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.
Barnor
Jacob Samson
JS
Department of Life and Environmental Science, Chiba Institute of Technology, Chiba, Japan.
Miyano-Kurosaki
Naoko
N
Yamaguchi
Kazuya
K
Abumi
Yusuke
Y
Ishikawa
Koichi
K
Yamamoto
Naoki
N
D000163:Acquired Immunodeficiency Syndrome / Q000628:therapy; D019380:Anti-HIV Agents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D015967:Gene Expression Regulation, Viral; D016346:Gene Products, vif / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D016325:HIV Long Terminal Repeat / Q000235:genetics*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D016086:Lentivirus / Q000235:genetics*; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D013997:Time Factors; D054320:vif Gene Products, Human Immunodeficiency Virus
D019380:Anti-HIV Agents; D016346:Gene Products, vif; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D054320:vif Gene Products, Human Immunodeficiency Virus