RNA Interference

Volume 623 of the series Methods in Molecular Biology pp 123-136


Strategies in Designing Multigene Expression Units to Downregulate HIV-1

  • Jane ZhangAffiliated withBeckman Research Institute of City of Hope Email author 
  • , John J. RossiAffiliated withBeckman Research Institute of City of Hope

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The treatment of viral diseases such as HIV and HCV is limited by the genetic diversity of the viruses, especially when they are under the selective pressure of drugs. This problem holds true for gene-based therapies using RNAi in which there is evolution of drug-resistant strains under the discriminating pressure of treatment (1, 2). In a gene therapy setting for treatment of HIV, the incorporation of multiple effector molecules, targeting different viral and cellular sequences, can improve viral inhibition by substantially delaying the emergence of escape mutants (3-8). However, for short hairpin RNA triggers of RNAi, high levels of expression by strong Pol III promoters has led to cell toxicity, and even death in experimental animals (9, 10). Here, we describe a new combinatorial anti-HIV gene expression system allowing simultaneous expression of multiple RNAi effector units from a single Pol II polycistronic transcript. Our platform is suitable for the inclusion of any shRNA sequence and can be combined with other types of small RNA antiviral inhibitors.

Key words

RNAi HIV-1 miRNAs siRNAs Polycistron