title: Unexpected origins of the enhanced pairing affinity of 2'-fluoro-modified RNA.
abstract: Various chemical modifications are currently being evaluated for improving the efficacy of short interfering RNA (siRNA) duplexes as antisense agents for gene silencing in vivo. Among the 2'-ribose modifications assessed to date, 2'deoxy-2'-fluoro-RNA (2'-F-RNA) has unique properties for RNA interference (RNAi) applications. Thus, 2'-F-modified nucleotides are well tolerated in the guide (antisense) and passenger (sense) siRNA strands and the corresponding duplexes lack immunostimulatory effects, enhance nuclease resistance and display improved efficacy in vitro and in vivo compared with unmodified siRNAs. To identify potential origins of the distinct behaviors of RNA and 2'-F-RNA we carried out thermodynamic and X-ray crystallographic analyses of fully and partially 2'-F-modified RNAs. Surprisingly, we found that the increased pairing affinity of 2'-F-RNA relative to RNA is not, as commonly assumed, the result of a favorable entropic contribution ('conformational preorganization'), but instead primarily based on enthalpy. Crystal structures at high resolution and osmotic stress demonstrate that the 2'-F-RNA duplex is less hydrated than the RNA duplex. The enthalpy-driven, higher stability of the former hints at the possibility that the 2'-substituent, in addition to its important function in sculpting RNA conformation, plays an underappreciated role in modulating Watson-Crick base pairing strength and potentially π-π stacking interactions.
authors.affiliation: Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, TN 37232, USA.
authors.forename: Pradeep S
authors.initials: PS
authors.lastname: Pallan
authors.forename: Emily M
authors.initials: EM
authors.lastname: Greene
authors.forename: Paul Andrei
authors.initials: PA
authors.lastname: Jicman
authors.forename: Rajendra K
authors.initials: RK
authors.lastname: Pandey
authors.forename: Muthiah
authors.initials: M
authors.lastname: Manoharan
authors.forename: Eriks
authors.initials: E
authors.lastname: Rozners
authors.forename: Martin
authors.initials: M
authors.lastname: Egli
mesh_terms: D018360:Crystallography, X-Ray; D005461:Fluorine / Q000737:chemistry*; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009997:Osmotic Pressure; D020871:RNA Stability; D012330:RNA, Double-Stranded / Q000737:chemistry*; D013816:Thermodynamics; D014867:Water / Q000737:chemistry
chemical_list: D012330:RNA, Double-Stranded; D014867:Water; D005461:Fluorine