This project provides support and enabling methodology for a multi-investigator effort to develop transcriptional gene silencing and activation as a new RNA-based strategy for controlling the infectivity of HIV-1. We will take advantage of exciting preliminary results that show that protein nanoparticles derived from the Q[bacteriophage can be chemically modified to carry multiple copies of the active RNA molecules, to target cells by virtue of a specifically engineered ligand-receptor interaction, and to deliver the active RNA to elicit the desired functional result. Many aspects of this platform technology remain to be optimized;such experiments and strategies are proposed here. They include the further use of unnatural amino acids in the nanoparticle structure to allow controlled chemical conjugation, the introduction of cleavable linkers to allow for RNA release, the introduction of polycationic species for charge compensation, the exploration of additional targeting ligands, and the cellular effects of all of these modifications. This work represents an attempt to address a longstanding challenge in oligonucleotide delivery, one that is extremely difficult to master on the scale required to implement RNA interference in vivo. However, the characteristics of transcriptional gene silencing/activation make it necessary to deliver smaller amounts of active agents, and thus these studies seek to further improve and study a promising protein nanoparticle delivery approach.
This project will develop and characterize the activity of protein nanoparticle platforms for the targeted delivery of active noncoding RNA molecules to cells that can be infected by, and can harbor in a latent state, the HIV-1 pathogen .
|Zhou, Jiehua; Lazar, Daniel; Li, Haitang et al. (2018) Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1. Theranostics 8:1575-1590|
|Astakhova, Kira; Ray, Roslyn; Taskova, Maria et al. (2018) ""Clicking"" Gene Therapeutics: A Successful Union of Chemistry and Biomedicine for New Solutions. Mol Pharm 15:2892-2899|
|Shevchenko, Galina; Morris, Kevin V (2018) All I's on the RADAR: role of ADAR in gene regulation. FEBS Lett 592:2860-2873|
|Shrivastava, Surya; Charlins, Paige; Ackley, Amanda et al. (2018) Stable Transcriptional Repression and Parasitism of HIV-1. Mol Ther Nucleic Acids 12:12-18|
|Johnsson, Per; Lister, Nicholas; Shevchenko, Galina et al. (2017) Reply to Liu et al.: Yin and yang of PTEN regulation. Proc Natl Acad Sci U S A 114:E10512-E10513|
|Lister, Nicholas; Shevchenko, Galina; Walshe, James L et al. (2017) The molecular dynamics of long noncoding RNA control of transcription in PTEN and its pseudogene. Proc Natl Acad Sci U S A 114:9942-9947|
|Hewson, Chris; Morris, Kevin V (2016) Form and Function of Exosome-Associated Long Non-coding RNAs in Cancer. Curr Top Microbiol Immunol 394:41-56|
|Lazar, Daniel C; Morris, Kevin V; Saayman, Sheena M (2016) The emerging role of long non-coding RNAs in HIV infection. Virus Res 212:114-26|
|Saayman, Sheena M; Lazar, Daniel C; Scott, Tristan A et al. (2016) Potent and Targeted Activation of Latent HIV-1 Using the CRISPR/dCas9 Activator Complex. Mol Ther 24:488-98|
|Weinberg, Marc S; Morris, Kevin V (2016) Transcriptional gene silencing in humans. Nucleic Acids Res 44:6505-17|
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