Sergei M. Gryaznov

Synthesis and properties of oligonucleotides containing aminodeoxythymidine units

Procedures are described for synthesis via solid support methodology of oligonucleotide analogues derived in part from 3-amino-3-deoxythymidine or 5-amino-5-deoxythymidine. Oligothymidylate decamers terminated with a 3-amino group or containing a 3-NHP(O)(O-)O-5 internucleoside link are found to form unusually stable complexes with poly(dA), poly(A), and oligo(dA). For related derivatives of 5-amino-5-deoxythymidine enhancement is less or absent, and in the case of multiple substitution destabilization of the heteroduplex may be observed. That the effect of the 3-amino group is general for oligonucleotide derivatives is indicated by enhanced Tm values for heteroduplex complexes of the mixed-base oligomer, d(TATTCAGTCAT(NH2)), and the methyl phosphonate derivatives, TmTmTmTmTmTmTmTmTmT(NH2) and d(TmAmTmTmCmAmGmTmCmAmT(NH2)).

A new approach to synthesis of Oligonucleotides with 3′ phosphoryl groups

A convenient method for synthesizing oligonucleotide 3′phosphates utilizes a diester phosphoramidate linker obtained by considering a nucleoside phosphoramidite reagent with an aminoalkyl solid support. Oligomer release is effected by NH3;H+.

Anchor for one step release of 3′-aminooligonucleotides from a solid support

Abstract A linker is described that permits covalently bound aminooligonucleotides to be liberated from a solid support by one step treatment with ammonium hydroxide.

Construction of branched DNA (bDNA) molecules by chemical ligation

Abstract Chemical ligation has been investigated in the assembly of branched DNA (bDNA) molecules. Water soluble coupling reagents were used to ligate single-stranded linear oligomers to bDNA molecules containing 5 and 15 branches. The ligations were particularly efficient when one of the oligomers was partially double-stranded constructed by branching to create a terminal ligation linker (AutoLink). Branched DNA molecules containing more than 1000 nucleotides were assembled.