2,2,5,5-tetramethylpyrrolidin-3-one-1-sulfinyl group for 5'-hydroxyl protection of deoxyribonucleoside phosphoramidites in the solid-phase preparation of DNA oligonucleotides.
Several nitrogen-sulfur reagents have been investigated as potential 5'-hydroxyl protecting groups for deoxyribonucleoside phosphoramidites to improve the synthesis of oligonucleotides on glass microarrays. Out of the nitrogen-sulfur-based protecting groups so far investigated, the 2,2,5,5-tetramethylpyrrolidin-3-one-1-sulfinyl group exhibited near optimal properties for 5'-hydroxyl protection by virtue of the mildness of its deprotection conditions. Specifically, the iterative cleavage of a terminal 5'-sulfamidite group in the synthesis of 5'-d(ATCCGTAGCCAAGGTCATGT) on controlled-pore glass is efficiently accomplished by treatment with iodine in the presence of an acidic salt. Hydrolysis of the oligonucleotide to its 2'-deoxyribonucleosides upon exposure to snake venom phosphodiesterase and bacterial alkaline phosphatase did not reveal the formation of any nucleobase adducts or other modifications. These findings indicate that the 2,2,5,5-tetramethylpyrrolidin-3-one-1-sulfinyl group for 5'-hydroxyl protection of phosphoramidites, such as 10a-d, may lead to the production of oligonucleotide microarrays exhibiting enhanced specificity and sensitivity in the detection of nucleic acid targets.
Division of Therapeutic Proteins, Center for Drug Evalution and Research, Food and Drug Administration, 8800 Rockville Pike, Bethesda, Maryland 20892, USA.
Vicente
V
Marchán
Jacek
J
Cieślak
Victor
V
Livengood
Serge L
SL
Beaucage
D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000138:chemical synthesis*; D003853:Deoxyribonucleosides / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000138:chemical synthesis*; D009943:Organophosphorus Compounds / Q000737:chemistry*; D011760:Pyrrolidinones / Q000737:chemistry*; D013452:Sulfonium Compounds / Q000737:chemistry*
D003853:Deoxyribonucleosides; D009841:Oligonucleotides; D009943:Organophosphorus Compounds; D011760:Pyrrolidinones; D013452:Sulfonium Compounds; C434331:phosphoramidite; D004247:DNA