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.

Public Health Relevance

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 .

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
1P01AI099783-01
Application #
8311228
Study Section
Special Emphasis Panel (ZAI1-RB-A (J1))
Project Start
Project End
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
1
Fiscal Year
2012
Total Cost
$113,825
Indirect Cost
$35,274
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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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
Fortes, Puri; Morris, Kevin V (2016) Long noncoding RNAs in viral infections. Virus Res 212:1-11
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
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
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Roberts, Thomas C; Hart, Jonathan R; Kaikkonen, Minna U et al. (2015) Quantification of nascent transcription by bromouridine immunocapture nuclear run-on RT-qPCR. Nat Protoc 10:1198-211
Yu, Albert D; Wang, Zichen; Morris, Kevin V (2015) Long noncoding RNAs: a potent source of regulation in immunity and disease. Immunol Cell Biol 93:277-83
Zhou, Jiehua; Satheesan, Sangeetha; Li, Haitang et al. (2015) Cell-specific RNA aptamer against human CCR5 specifically targets HIV-1 susceptible cells and inhibits HIV-1 infectivity. Chem Biol 22:379-90
Vadie, Nadia; Saayman, Sheena; Lenox, Alexandra et al. (2015) MYCNOS functions as an antisense RNA regulating MYCN. RNA Biol 12:893-9

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