The efficacy of siRNAs against hepatitis C virus is strongly influenced by structure and target site accessibility.
Hepatitis C virus (HCV) is a global health problem. Designing therapeutic agents that target HCV's RNA genome remains challenging. HCV genomic RNA is large and highly structured with long-range genome-scale ordered RNA structures. Predicting the secondary- and tertiary-structure elements that reveal the accessibility of target sites within HCV RNA is difficult because of the abundance of long-range interactions. Target site accessibility remains a significant barrier to the design of effective therapeutics such as small interfering RNAs (siRNAs) against different strains of HCV. Here we developed two methods that interrogate the folding of HCV RNA, an approach involving viral RNA microarrays (VRMs) and an HCV viral RNA-coated magnetic bead-based (VRB) assay. VRMs and VRBs were used to determine target site accessibility for siRNAs designed against the HCV genome. Both methods predicted potency of siRNAs in cell-culture models for HCV replication that are not easily predicted by informatic means.
Sagan
Selena M
SM
Steacie Institute for Molecular Sciences, The National Research Council of Canada, Ottawa, ON K1A 0R6, Canada. smsagan@stanford.edu
Nasheri
Neda
N
Luebbert
Christian
C
Pezacki
John Paul
JP
D045744:Cell Line, Tumor; D055785:Gene Knockdown Techniques; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D008280:Magnetics; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry*; D014779:Virus Replication / Q000187:drug effects
D034741:RNA, Small Interfering; D012367:RNA, Viral