title: Acetal levulinyl ester (ALE) groups for 2'-hydroxyl protection of ribonucleosides in the synthesis of oligoribonucleotides on glass and microarrays.
abstract: We describe a synthetic strategy that permits both the growth and deprotection of RNA chains that remain attached to a solid polymer support or chip surface. The key synthons for RNA synthesis are novel 5'-O-DMTr 2'-acetal levulinyl ester (2'-O-ALE) ribonucleoside 3'-phosphoramidite derivatives. In the presence of 4,5-dicyanoimidazole (DCI) as the activator, these monomers coupled to Q-CPG solid support with excellent coupling efficiency (approximately 98.7%). The method was extended to the light directed synthesis of poly rU and poly rA on a microarray through the use of a 5'-O-(2-(2-nitrophenyl)propoxycarbonyl)-2'-O-ALE-3'-phosphoramidite derivative. A two-stage deprotection strategy was employed to fully deblock the RNA directly on the Q-CPG or microarray support without releasing it from the support's surface: phosphate group deblocking with NEt(3) in acetonitrile (ACN) (2:3 v/v; 1 h, r.t.) followed by removal of the 2'-O-ALE groups under mild hydrazinolysis conditions (0.5-4 h, r.t.). This last treatment also removed the levulinyl (Lv) group on adenine (N(6)) and cytosine (N(4)) and the dimethylformamidine (dmf) group on guanine (N(2)). The chemistry and methods described here pave the way to the fabrication of microarrays of immobilized RNA probes for analyzing molecular interactions of biological interest.
authors.affiliation: Department of Chemistry, McGill University, Montreal, Quebec, Canada H3A 2K6.
authors.forename: Jeremy G
authors.initials: JG
authors.lastname: Lackey
authors.forename: Debbie
authors.initials: D
authors.lastname: Mitra
authors.forename: Mark M
authors.initials: MM
authors.lastname: Somoza
authors.forename: Franco
authors.initials: F
authors.lastname: Cerrina
authors.forename: Masad J
authors.initials: MJ
authors.lastname: Damha
mesh_terms: D000080:Acetals / Q000737:chemistry; D004952:Esters / Q000737:chemistry; D007982:Levulinic Acids / Q000737:chemistry; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009843:Oligoribonucleotides / Q000138:chemical synthesis*; D009943:Organophosphorus Compounds / Q000737:chemistry; D016372:RNA, Antisense / Q000737:chemistry; D012263:Ribonucleosides / Q000737:chemistry*
chemical_list: D000080:Acetals; D004952:Esters; D007982:Levulinic Acids; D009843:Oligoribonucleotides; D009943:Organophosphorus Compounds; D016372:RNA, Antisense; D012263:Ribonucleosides; C434331:phosphoramidite