Intracellular inhibition of HIV-1 replication using a dual protein- and RNA-based strategy

Abstract

Exporting unspliced human immunodeficiency virus type 1 function, thereby inhibiting viral replication. In the present (HIV-1) RNA from the nucleus to the cytoplasm, through studies, different HIV-1 RRE region-specific hammerhead an interaction between the viral regulatory Rev protein and ribozymes were constructed and their anti-HIV-1 repliRev response element (RRE) RNA, is a critical step in the cation effects were assayed in diverse RNA polymerase HIV-1 life-cycle. Disruption of either Rev or the RRE will (pol) II and III promoters and vector systems in cell culture. completely inhibit HIV-1 replication. As such, a strategy for Utilizing this combination of an SFv and a ribozyme as a somatic gene therapy to treat HIV-1 infection by intracelludual strategy to block HIV-1 replication, both at the protein lar expression of an anti-HIV-1 Rev single chain variable and RNA level, data from these studies demonstrated that fragment (SFv) and a ribozyme which specifically targets potent inhibition of HIV-1 replication can be achieved via the RRE was developed. The anti-Rev D8SFv, which this approach. Combination gene therapies hold promise, specifically targets the Rev activation domain, may be a analogous to combination chemotherapeutic regimens, for key component of combination intracellular immunization, the in vivo treatment of HIV-1 infections. as it has been previously shown to potently inhibit Rev

Functional Neutralization of HIV-1 Vif Protein by Intracellular Immunization Inhibits Reverse Transcription and Viral Replication

“…The results presented here extend previous studies demonstrating that high affinity single-chain antibodies can be stably expressed in the cytoplasm (31,34,36,35,37). Folding of the scFv in the reducing environment of the cytoplasm can form functional binding sites as demonstrated by co-immunoprecipitation.…”

“…Intrabodies are single-chain variable region antibody fragments expressed and confined intracellularly, where they can bind to viral proteins and other targets (29,31,(33)(34)(35)(36)(37). The early events of the viral life cycle are potential therapeutic targets that could be inhibited using anti-HIV-1 Vif intrabodybased strategies and may therefore represent a new therapeutic approach to inhibit HIV-1 reverse transcription.…”

Hammerhead ribozyme design and application

“…Cotransfection of HIV-1 DNA and several multimeric ribozyme expression vectors into permissive cell lines resulted in a specific inhibition of HIV-1 replication, indicating that the multitarget ribozymes were also functional intracellularly. A variation of the multitarget HIV-1-specific ribozyme has been described in which single-target or multiple-target ribozyme genes are transcribed in fusion with decoy sequences for sequestration of transactivating proteins or binding specific secondary structure elements (like the stem-loop II of the Rev response element) on the target RNA recognized by such proteins [149,159,160,[182][183][184]. The decoys thus function by competing with either virus RNA or transactivating protein to inhibit their association.…”

Gene therapy for HIV-1

“…Interfering RNA-and protein-based strategies may also be combined. Antisense or sense RNAs were combined with dominant negative proteins or intrabodies [42,43] showing to confer better protection than the single anti-HIV genes.…”