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Conceptual “Heat‐Driven” Approach to the Synthesis of DNA Oligonucleotides on Microarrays
Author(s) -
GRAJKOWSKI A.,
CIEŚLAK J.,
CHMIELEWSKI M. K.,
MARCHÁN V.,
PHILLIPS L. R.,
WILK A.,
BEAUCAGE S. L.
Publication year - 2003
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1281.003
Subject(s) - oligonucleotide , deoxyribonucleoside , phosphoramidite , chemistry , nucleobase , oligonucleotide synthesis , monomer , protecting group , combinatorial chemistry , solid phase synthesis , dna , nucleotide , dna synthesis , phosphate , dna microarray , stereochemistry , biochemistry , organic chemistry , polymer , peptide , alkyl , gene expression , gene
A bstract : The discovery of deoxyribonucleoside cyclic N ‐acylphosphoramidites, a novel class of phosphoramidite monomers for solid‐phase oligonucleotide synthesis, has led to the development of a number of phosphate protecting groups that can be cleaved from DNA oligonucleotides under thermolytic neutral conditions. These include the 2‐( N ‐formyl‐ N ‐methyl)aminoethyl, 4‐oxopentyl, 3‐( N‐tert ‐butyl)carboxamido‐1‐propyl, 3‐(2‐pyridyl)‐1‐propyl, 2‐[ N ‐methyl‐ N ‐(2‐pyridyl)]aminoethyl, and 4‐methythiobutyl groups. When used for 5′‐hydroxyl protection of nucleosides, the analogous 1‐phenyl‐2‐[ N ‐methyl‐ N ‐(2‐pyridyl)]aminoethyloxycarbonyl group exhibited excellent thermolytic properties, which may permit an iterative “heat‐driven” synthesis of DNA oligonucleotides on microarrays. In this regard, progress has been made toward the use of deoxyribonucleoside cyclic N ‐acylphosphoramidites in solid‐phase oligonucleotide syntheses without nucleobase protection. Given that deoxyribonucleoside cyclic N ‐acylphosphoramidites produce oligonucleotides with heat‐sensitive phosphate protecting groups, blocking the 5′‐hydroxyl of these monomers with, for example, the thermolabile 1‐phenyl‐2‐[ N ‐methyl‐ N ‐(2‐pyridyl)]aminoethyloxycarbonyl group may provide a convenient thermo‐controlled method for the synthesis of oligonucleotides on microarrays.