In Vivo GFP Knockdown by Cationic Nanogel-siRNA Polyplexes

Bioengineering

Arun R. Shrivats²  Yuji Mishina¹  Saadyah Averick³  Krzysztof Matyjaszewski³  Jeffrey O. Hollinger² 

[1] School of Dentistry, University of Michigan, 1011 N. University Ave., Ann Arbor, MI 48109, USA; E-Mail:;Department of Biomedical Engineering, Carnegie Mellon University, 700 Technology Dr., Pittsburgh, PA 15219, USA; E-Mail:;Department of Chemistry, Carnegie Mellon University, 4400 Fifth Ave., Pittsburgh, PA 15213, USA; E-Mails:

关键词: gene therapy;    gene expression;    RNA Interference;    siRNA;    GFP;    ATRP;    nanostructured polymer;   

DOI  :  10.3390/bioengineering2030160

来源: mdpi

【 摘 要 】

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) for in vitro and in vivo siRNA 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 against Gapdh significantly reduced GAPDH enzyme activity in mammalian cell culture models supplemented with 10% serum. Moreover, nanogel NSP-mediated siRNA delivery significantly inhibited in vivo GFP 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.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.