A new and versatile reagent for incorporating multiple primary aliphatic amines into synthetic oligonucleotides

Abstract

A novel and versatile phosphoramidite, N-Fmoc-O1-DMT-O2-cyanoethoxydiisopropylamino-phosphinyl-3-am ino-1,2-propanediol (1, Fig. 1), has been synthesized and used to incorporate primary aliphatic amines into synthetic oligonucleotides. Its convenient preparation and use in solid phase oligonucleotide synthesis is described. Using phosphoramidite 1, an amino-modified oligonucleotide probe complementary to M13mp18 DNA was constructed with five primary amines attached to the 5'-terminus. The amino-modified oligonucleotide was subsequently labeled with biotin and employed in a dot-blot hybridization assay. As little as 0.5 ng of M13mp18 target DNA was colorimetrically detected.

Triplex formation inhibits HER-2/neu transcription in vitro.

“…It would be desirable to bridge these interruptions in a manner that minimizes steric interactions that could be destabilizing to triplex formation. An abasic molecule that maintains the proper internucleotide spacing (27) would be ideally suited as such a bridge. We have examined the effect ofsubstituting an abasic bridge for a pyrimidine residue in a purine-rich triplex forming oligonucleotide.…”

Synthesis and antibody-mediated detection of oligonucleotides containing multiple 2,4-dinitrophenyl reporter groups

“…(ii) An amino group separated from the backbone by a linker to allow attachment of the DNP group. Misiura et al (1) made use of a 3-carbon glycerol type backbone in the construction of a biotin phosphoramidite, while Nelson et al (27) have used the readily available 3-amino-1,2-propanediol unit to introduce multiple primary amino groups to the 5' end of oligonucleotides. We describe here the synthesis of a range of DNP monomers involving the use of these backbones, both in their basic forms, and in molecules which involve extension of their amino functionalities to form a linker arm.…”

Development and Applications of Fluorescent Oligonucleotides

“…The same functional groups can also be incorporated at the 3'-end of the ONs via modified supports [135]. Post-synthetic coupling of labels at either internucleotidic positions via insertion of nonnatural linkages [145] or nucleic bases has also been reported [146][147][148].…”