Efficient assessment of modified nucleoside stability under conditions of automated oligonucleotide synthesis: characterization of the oxidation and oxidative desulfurization of 2-thiouridine.
In order to efficiently assess the chemical stability of modified nucleosides to the reagents and conditions of automated oligonucleotide synthesis, we designed, developed and tested a scheme in which the modified nucleoside, directly attached to a solid support, is exposed to the cyclic chemistry of the instrument. Stability of 2-thiouridine against different oxidizers was investigated. Tertbutyl hydroperoxide (1 M) in anhydrous acetonitrile was a more effective oxidizer for the incorporation of 2-thiouridine into oligonucleotide chains than the same oxidizer in methylene chloride. Carbon tetrachloride/water in the presence of a basic catalyst was superior in maintaining the thiocarbonyl function, but its utility for RNA synthesis has yet to be fully tested, whereas 2-phenylsulfonyloxaziridine was a very efficient reagent for oxidative desulfurization of 2-thiouridine.
Sochacka
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Institute of Organic Chemistry, Technical University of Lódź, Poland.
D000097:Acetonitriles / Q000737:chemistry; D002251:Carbon Tetrachloride / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D006861:Hydrogen Peroxide / Q000737:chemistry; D008956:Models, Chemical; D009691:Nucleic Acid Denaturation; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry*; D010100:Oxygen / Q000378:metabolism*; D013455:Sulfur / Q000737:chemistry*; D013891:Thiouridine / Q000031:analogs & derivatives* / Q000737:chemistry*; D013997:Time Factors
C105273:2-thiouridine; D000097:Acetonitriles; D009841:Oligonucleotides; D013891:Thiouridine; D013455:Sulfur; D006861:Hydrogen Peroxide; D002251:Carbon Tetrachloride; D010100:Oxygen; C032159:acetonitrile