title: Preparation of Intrastrand {G}O(6) -Alkylene-O(6) {G} Cross-Linked Oligonucleotides.
abstract: This unit describes the preparation O(6) -2'-deoxyguanosine-butylene-O(6) -2'-deoxyguanosine dimer phosphoramidites and precursors for incorporation of site-specific intrastrand cross-links (IaCL) into DNA oligonucleotides. Protected 2'-deoxyguanosine dimers are produced using the Mitsunobu reaction. IaCL DNA containing the intradimer phosphodiester are first chemically phosphorylated, followed by a ring-closing reaction using the condensing reagent 1-(2-mesitylenesulfonyl)-3-nitro-1H-1,2,4-triazole. Phosphoramidites are incorporated into oligonucleotides by solid-phase synthesis and standard deprotection and cleavage protocols are employed. This approach allows for the preparation of IaCL DNA substrates in amounts and purity amenable for biophysical characterization, and biochemical studies as substrates to investigate DNA repair and bypass pathways. © 2016 by John Wiley & Sons, Inc.
authors: Department of Chemistry and Biochemistry, Concordia University, Montréal, Québec, Canada.
authors: Derek K
authors: DK
authors: O'Flaherty
authors: Department of Chemistry and Biochemistry, Concordia University, Montréal, Québec, Canada.
authors: Christopher J
authors: CJ
authors: Wilds
mesh_terms: D003432:Cross-Linking Reagents / Q000737:chemistry; D003849:Deoxyguanosine / Q000737:chemistry; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000737:chemistry*; D060327:Solid-Phase Synthesis Techniques
chemical_list: D003432:Cross-Linking Reagents; D009841:Oligonucleotides; D009943:Organophosphorus Compounds; C434331:phosphoramidite; D003849:Deoxyguanosine
keywords: DNA damage; DNA intrastrand cross-link; DNA repair and bypass; chemically modified oligonucleotide; oligonucleotide synthesis; solid-phase synthesis