In VivoGFP Knockdown by Cationic Nanogel-siRNA Polyplexes.
RNA interference (RNAi) is a powerful tool to treat diseases and elucidate target gene function. Prior to clinical implementation, however, challenges including the safe, efficient and targeted delivery of siRNA must be addressed. Here, we report cationic nanogel nanostructured polymers (NSPs) prepared by atom transfer radical polymerization (ATRP) forin vitroandin vivosiRNA delivery in mammalian models. Outcomes from siRNA protection studies suggested that nanogel NSPs reduce enzymatic degradation of siRNA within polyplexes. Further, the methylation of siRNA may enhance nuclease resistance without compromising gene knockdown potency. NSP-mediated RNAi treatments againstGapdhsignificantly reduced GAPDH enzyme activity in mammalian cell culture models supplemented with 10% serum. Moreover, nanogel NSP-mediated siRNA delivery significantly inhibitedin vivoGFP expression in a mouse model. GFP knockdown was siRNA sequence-dependent and facilitated by nanogel NSP carriers. Continued testing of NSP/siRNA compositions in disease models may produce important new therapeutic options for patient care.
Department of Biomedical Engineering, Carnegie Mellon University, 700 Technology Dr., Pittsburgh, PA 15219, USA.
Arun R
AR
Shrivats
School of Dentistry, University of Michigan, 1011 N. University Ave., Ann Arbor, MI 48109, USA.
Yuji
Y
Mishina
Department of Chemistry, Carnegie Mellon University, 4400 Fifth Ave., Pittsburgh, PA 15213, USA.
Saadyah
S
Averick
Department of Chemistry, Carnegie Mellon University, 4400 Fifth Ave., Pittsburgh, PA 15213, USA.
Krzysztof
K
Matyjaszewski
Department of Biomedical Engineering, Carnegie Mellon University, 700 Technology Dr., Pittsburgh, PA 15219, USA.
Jeffrey O
JO
Hollinger
ATRP; GFP; RNA Interference; gene expression; gene therapy; nanostructured polymer; siRNA