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 .
|Johnsson, Per; Lipovich, Leonard; Grander, Dan et al. (2014) Evolutionary conservation of long non-coding RNAs; sequence, structure, function. Biochim Biophys Acta 1840:1063-71|
|Damski, Caio; Morris, Kevin V (2014) Targeted small noncoding RNA-directed gene activation in human cells. Methods Mol Biol 1173:1-10|
|Zhou, Jiehua; Rossi, John (2014) Cell-type-specific aptamer and aptamer-small interfering RNA conjugates for targeted human immunodeficiency virus type 1 therapy. J Investig Med 62:914-9|
|Saayman, Sheena; Ackley, Amanda; Turner, Anne-Marie W et al. (2014) An HIV-encoded antisense long noncoding RNA epigenetically regulates viral transcription. Mol Ther 22:1164-75|
|Groen, Jessica N; Capraro, David; Morris, Kevin V (2014) The emerging role of pseudogene expressed non-coding RNAs in cellular functions. Int J Biochem Cell Biol 54:350-5|
|Johnsson, Per; Morris, Kevin V; Grandér, Dan (2014) Pseudogenes: a novel source of trans-acting antisense RNAs. Methods Mol Biol 1167:213-26|
|Morris, Kevin V; Mattick, John S (2014) The rise of regulatory RNA. Nat Rev Genet 15:423-37|
|Roberts, Thomas C; Morris, Kevin V; Weinberg, Marc S (2014) Perspectives on the mechanism of transcriptional regulation by long non-coding RNAs. Epigenetics 9:13-20|
|Groen, Jessica N; Morris, Kevin V (2013) Chromatin, non-coding RNAs, and the expression of HIV. Viruses 5:1633-45|
|Akkina, Ramesh (2013) Human immune responses and potential for vaccine assessment in humanized mice. Curr Opin Immunol 25:403-9|
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