title: Nucleoside optimization for RNAi: a high-throughput platform.
abstract: The RNA induced silencing complex (RISC) contains at its core the endonuclease Argonaute (Ago) that allows for guide strand (GS)-mediated sequence-specific cleavage of the target mRNA. Functionalization of the sugar/phosphodiester backbone of the GS, which is in direct contact with Ago, presents a logical opportunity to affect RISC's activity. A systematic evaluation of modified nucleosides requires the synthesis of phosphoramidites corresponding to all four canonical bases (A, U, C, and G) and their sequential evaluation at each position along the 21-nucleotide-long GS. With the use of a platform approach, the sequential replacement of canonical bases with inosine greatly simplifies the problem and defines a new activity baseline toward which the corresponding sugar-modified inosines are compared. This approach was validated using 2'-O-benzyl modification, which demonstrated that positions 5, 8, 15, and 19 can accommodate this large group. Application of this high-throughput methodology now allows for hypothesis-driven rational design of highly potent, immunologically silent and stable siRNAs suitable for therapeutic applications.
authors.affiliation: Department of Process Chemistry, Merck & Co., Rahway, New Jersey 07065, USA. gabor_butora@merck.com
authors.forename: Gabor
authors.initials: G
authors.lastname: Butora
authors.forename: Denise M
authors.initials: DM
authors.lastname: Kenski
authors.forename: Abby J
authors.initials: AJ
authors.lastname: Cooper
authors.forename: Wenlang
authors.initials: W
authors.lastname: Fu
authors.forename: Ning
authors.initials: N
authors.lastname: Qi
authors.forename: Jenny J
authors.initials: JJ
authors.lastname: Li
authors.forename: W Michael
authors.initials: WM
authors.lastname: Flanagan
authors.forename: Ian W
authors.initials: IW
authors.lastname: Davies
mesh_terms: D001483:Base Sequence; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009705:Nucleosides / Q000737:chemistry* / Q000235:genetics; D034622:RNA Interference
chemical_list: D009705:Nucleosides