Quantitative evaluation of siRNA delivery in vivo

• DOI: 10.1261/rna.2255810
• Publication Date: 2010-10-13T07:01:56Z
• Authors: Yi Pei; Paula J. Hancock; Hangchun Zhang; René Bartz; Craig Cherrin; Nathalie Innocent; Colin J. Pomerantz; Jessica Seitzer; Martin L. Koser; Marc T. Abrams; Yan Xu; Nelly A. Kuklin; Paul A. Burke; Alan B. Sachs; Laura Sepp-Lorenzino; Stanley F. Barnett
• Keywords: Trans-acting siRNA; Argonaute

Effective small interfering RNA (siRNA)–mediated therapeutics require the siRNA to be delivered into cellular RNA-induced silencing complex (RISC). Quantitative information of this essential delivery step is currently inferred from efficacy gene and uptake in tissue. Here we report an approach directly quantify RISC rodents monkey. This achieved by specific immunoprecipitation tissue lysates quantification RNAs immunoprecipitates stem–loop PCR. The method, expected independent vehicle target, label-free, throughput acceptable for preclinical animal studies. We characterized a lipid-formulated integrating these approaches obtained quantitative perspective on accumulation, loading, silencing. described methodologies have utility study mechanism, development therapeutics, clinical trial design.