Evaluation of the Efficacy And Toxicity of RNAs Targeting HIV-1 Production for Use in Gene or Drug Therapy.
Small RNA therapies targeting post-integration steps in the HIV-1 replication cycle are among the top candidates for gene therapy and have the potential to be used as drug therapies for HIV-1 infection. Post-integration inhibitors include ribozymes, short hairpin (sh) RNAs, small interfering (si) RNAs, U1 interference (U1i) RNAs and RNA aptamers. Many of these have been identified using transient co-transfection assays with an HIV-1 expression plasmid and some have advanced to clinical trials. In addition to measures of efficacy, small RNAs have been evaluated for their potential to affect the expression of human RNAs, alter cell growth and/or differentiation, and elicit innate immune responses. In the protocols described here, a set of transient transfection assays designed to evaluate the efficacy and toxicity of RNA molecules targeting post-integration steps in the HIV-1 replication cycle are described. We have used these assays to identify new ribozymes and optimize the format of shRNAs and siRNAs targeting HIV-1 RNA. The methods provide a quick set of assays that are useful for screening new anti-HIV-1 RNAs and could be adapted to screen other post-integration inhibitors of HIV-1 replication.
Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research; Department of Microbiology & Immunology, McGill University.
Robert J
RJ
Scarborough
Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research; Department of Microbiology & Immunology, McGill University.
Kelsey L
KL
Adams
Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research; Department of Microbiology & Immunology, McGill University.
Olivier
O
Del Corpo
Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research.
Aïcha
A
Daher
Virus-Cell Interactions Laboratory, Lady Davis Institute for Medical Research; Department of Microbiology & Immunology, McGill University; Department of Medicine, McGill University; anne.gatignol@mcgill.ca.
Anne
A
Gatignol
D001681:Biological Assay / Q000379:methods; D004358:Drug Therapy; D015316:Genetic Therapy; D015658:HIV Infections / Q000188:drug therapy; D015497:HIV-1; D006801:Humans; D034622:RNA Interference; D016337:RNA, Catalytic; D034741:RNA, Small Interfering; D014162:Transfection; D014779:Virus Replication
D016337:RNA, Catalytic; D034741:RNA, Small Interfering
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