Identification of metabolically stable 5′-phosphate analogs that support single-stranded siRNA activity

作者：Thazha P. Prakash, Walt F. Lima, Heather Murray, Wenyu Li, Garth A. Kinberger, Alfred E. Chappell, Hans Gaus, Punit P. Seth, Balkrishen Bhat, Stanley T. Crooke, Eric E. Swayze · 发表于：Nucleic Acids Research · 年份：2015 · DOI：10.1093/nar/gkv162 · 被引用次数：96 · 研究领域：RNA Interference and Gene Delivery、Advanced biosensing and bioanalysis techniques、DNA and Nucleic Acid Chemistry

The ss-siRNA activity in vivo requires a metabolically stable 5'-phosphate analog. In this report we used crystal structure of the 5'-phosphate binding pocket of Ago-2 bound with guide strand to design and synthesize ss-siRNAs containing various 5'-phosphate analogs. Our results indicate that the electronic and spatial orientation of the 5'-phosphate analog was critical for ss-siRNA activity. Chemically modified ss-siRNA targeting human apoC III mRNA demonstrated good potency for inhibiting ApoC III mRNA and protein in transgenic mice. Moreover, ApoC III ss-siRNAs were able to reduce the triglyceride and LDL cholesterol in transgenic mice demonstrating pharmacological effect of ss-siRNA. Our study provides guidance to develop surrogate phosphate analog for ss-siRNA and demonstrates that ss-siRNA provides an alternative strategy for therapeutic gene silencing.