CRISPR-based RNA targeting for HIV intervention

Human Immunodeficiency Virus (HIV) persists in the form of a ???latent reservoir??? hiding from complete eradication by the immune system or antiretroviral drugs. Current drugs eliminate actively multiplying HIV but are unable to eradicate the latent virus. Therefore. we hypothesize that a successful HIV RNA targeting strategy will efficiently and simultaneously target multiple conserved sites in HIV transcript. the possibility of concurrent alterations to which will either be negligible or entail fitness cost to the virus. Our preliminary data showed differential expression of cellular long non-coding RNAs (LncRNA) in elite controllers. which show spontaneous. long-term control of HIV. Preliminary screen using the CRISPR/Cas13 mediated silencing of EC-associated lncRNAs showed inhibition of HIV replication. These findings lead us to hypothesize that significant suppression of specific lncRNAs in Elite controllers is protective against HIV. In this proposed project. we will pioneer the use of the differential expression of non-coding RNAs in elite controllers to understand the role of these non-coding RNAs in HIV replication. host resistance. and disease outcomes. We propose to optimize the use of this targeting strategy against HIV and delivery to the primary cells. In the proposed project. we will carry out the optimization and efficacy of Adeno- associated virus and nanoparticle-mediated in vivo delivery of lncRNA-silencing cargo in mice and confirm its delivery using advanced imaging techniques. This approach has the potential to mediate long- term expression of a corrective payload to achieve long term function cure of HIV.