title: Phototriggered DNA phosphoramidate ligation in a tandem 5'-amine deprotection/3'-imidazole activated phosphate coupling reaction.
abstract: We report the preparation and use of an N-methyl picolinium carbamate protecting group for applications in a phototriggered nonenzymatic DNA phosphoramidate ligation reaction. Selective 5'-amino protection of a modified 13-mer oligonucleotide is achieved in aqueous solution by reaction with an N-methyl-4-picolinium carbonyl imidazole triflate protecting group precursor. Deprotection is carried out by photoinduced electron transfer from Ru(bpy)(3)(2+) using visible light photolysis and ascorbic acid as a sacrificial electron donor. Phototriggered 5'- amino oligonucleotide deprotection is used to initiate a nonenzymatic ligation of the 13-mer to an imidazole activated 3'-phospho-hairpin template to generate a ligated product with a phosphoramidate linkage. We demonstrate that this methodology offers a simple way to exert control over reaction initiation and rates in nonenzymatic DNA ligation for potential applications in the study of model protocellular systems and prebiotic nucleic acid synthesis.
authors.affiliation: Material, Physics and Applications Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Cape
authors.forename: Joseph B
authors.initials: JB
authors.lastname: Edson
authors.forename: Liam P
authors.initials: LP
authors.lastname: Spencer
authors.forename: Michael S
authors.initials: MS
authors.lastname: DeClue
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Ziock
authors.forename: Sarah
authors.initials: S
authors.lastname: Maurer
authors.forename: Steen
authors.initials: S
authors.lastname: Rasmussen
authors.forename: Pierre-Alain
authors.initials: PA
authors.lastname: Monnard
authors.forename: James M
authors.initials: JM
authors.lastname: Boncella
mesh_terms: D000577:Amides / Q000737:chemistry*; D000588:Amines / Q000737:chemistry*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D007093:Imidazoles / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000235:genetics; D010710:Phosphates / Q000737:chemistry*; D010756:Phosphoric Acids / Q000737:chemistry*; D055668:Photochemical Processes
chemical_list: D000577:Amides; D000588:Amines; D007093:Imidazoles; D009838:Oligodeoxyribonucleotides; D010710:Phosphates; D010756:Phosphoric Acids; C029899:imidazole; D004247:DNA; C011067:phosphoramidic acid