Journal of Nanomedicine & Nanotechnology

Cobalt oxide nanoparticle mediated delivery of Splice Switching Oligonucleotides (SSO) RNA

8^th World Medical Nanotechnology Congress & Expo

June 08-09, 2016 Dallas, USA

Miranda N Hurst, Meghana Ramani, John M Dean, Parwathy Chandran, Nancy A Monteiro-Riviere and Robert K DeLong

Kansas State University, USA
Missouri State University, USA
Washington University, USA

Scientific Tracks Abstracts: J Nanomed Nanotechnol

Abstract:

Splice Switching Oligonucleotides (SSOs) are a class of antisense RNA that directly modulates pre-mRNA splicing. SSOs require a delivery vehicle to reach the nucleus such as nanoparticles. An assay has been developed using cancer cells stably transfected with luciferase reporter gene that is interrupted by human -globin intron. This construct results in production of non-functional luciferase protein. Upon successful nuclear delivery of 705 SSO, the intron is correctly spliced out resulting in expression of functional luciferase76. We analyzed nanoparticle delivery of SSO to cancer cells. We observed that Co3O4 (size < 100 nm) displayed the highest payload (83 nM SSO per mM nanoparticle). Binding constants were evaluated by UV-vis absorption (2.2 x 106 M-1) and fluorescence spectroscopies (2.128 x 106 M-1). Nanosight and DLS analyses indicate increase in NP size upon complexation. Positive zeta potential of Co3O4 shifts towards the negative indicating electrostatic interaction. First generation nanoparticles were screened for SSO delivery to A375 pLuc cells measuring photoluminescence. Cobalt oxide was found to deliver SSO in a dose-dependent manner. Cytotoxicity evaluation against HEK cell lines indicates 100% cell viability after 24 hrs of Co3O4 treatment. Significant increase in splicing correction as compared to the control was observed by RT-PCR and gel electrophoresis. Binding, uptake and delivery is being further evaluated by multiple microscopy methods. Overall, these results support our claim that unmodified Co3O4 nanoparticle mediated SSO delivery corrects RNA splicing and restores active Luciferase expression.

Biography :

Miranda N Hurst received her bachelor of science in cell and molecular biology from Missouri State University in 2015. She is currently working as a Research Assistant under Dr. Robert Delong at the Nanotechnology Innovation Center of Kansas State (NICKS). Her current research focuses on the stabilization and delivery of RNA via nanoparticles into human and mouse cell lines. Her research interests include the relationship between structure and function of RNA and protein in the complex cellular environment, specifically for the development of cancer therapies. She has two publications in peer reviewed journals and one patent submitted.

Email: mnhurst@ksu.edu