title: Preparation of 5'-silyl-2'-orthoester ribonucleosides for use in oligoribonucleotide synthesis.
abstract: The recent discovery that small interfering RNAs (siRNAs) induce gene suppression in mammalian cells has sparked tremendous interest in using siRNA-based assays and high-throughput screens to study gene function. As a result, research programs at leading academic and commercial institutions have become a substantial and rapidly growing market for synthetic RNA. Important considerations in synthesizing RNA for biological gene function studies are sequence integrity, purity, scalability, and resistance to nucleases; ease of chemical modification, deprotection, and handling; and cost. Of the well-established RNA synthesis methods, 2'-ACE chemistry is the only one that meets all of these criteria. 2'-ACE technology employs a unique class of silyl ethers to protect the 5'-hydroxyl, in combination with an acid-labile orthoester protecting group on the 2'-hydroxyl (2'-ACE). 2'-ACE-protected phosphoramidite monomers are joined using standard solid-phase technology to achieve RNA synthesis at efficiencies rivaling those for DNA. This unit describes the synthesis of standard 5'-silyl-2'-ACE-protected phosphoramidites.
authors.affiliation: Dharmacon Inc., Lafayette, Colorado, USA.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Scaringe
authors.forename: David
authors.initials: D
authors.lastname: Kitchen
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Kaiser
authors.forename: William S
authors.initials: WS
authors.lastname: Marshall
mesh_terms: D019411:Clinical Laboratory Techniques; D008954:Models, Biological; D009843:Oligoribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000138:chemical synthesis / Q000737:chemistry; D012263:Ribonucleosides / Q000138:chemical synthesis* / Q000737:chemistry*
chemical_list: D009843:Oligoribonucleotides; D009943:Organophosphorus Compounds; D012263:Ribonucleosides; C434331:phosphoramidite