US 5,683,985 A · Oligonucleotide decoys and methods relating thereto

Abstract

Improved DNA structures are disclosed which contain target sequences which bind to control proteins (such as the CREB protein). The structures of the present invention are stable to degradation, and are effective as decoys for control proteins, making it possible to modulate the transcriptional control normally exerted by such control proteins. In addition, there is provided a method to reversibly crosslink oligonucleotides to polypeptides which recognize the oligonucleotide sequence. This method involves synthesizing DNA structures as described above, wherein one or more phosphorothioate diester linkages are incorporated into the resulting oligonucleotide, allowing the phosphorothioate diester-containing oligonucleotide to bind to polypeptides which recognize the sequence of said oligonucleotide, then contacting the polypeptide-bound oligonucleotide with a transition metal reagent.

Claims20

Dependent claims

An oligonucleotide for modulating transcription activation of trans-acting factor responsive genes, said oligonucleotide comprising, reading from the 5'-end of said oligonucleotide a first segment, a second segment and a third segment, wherein:

(i) said first segment comprises a sequence of deoxynucleotide or nucleotide residues, said sequence comprising a first core recognition sequence having flanking bases on either side;
wherein said first segment, when hybridized with the third segment, forms at least one transcription control recognition sequence of at least 6 nucleotides,
(ii) said second segment comprises a sequence of deoxynucleotide or nucleotide residues, said sequence having a length sufficient to allow the formation of a first loop structure between the 3'-end of said first segment and the 5'-end of the third segment, and
(iii) said third segment comprises a sequence of deoxynucleotide or nucleotide residues, said sequence comprising a second core recognition sequence having flanking bases on either side, and wherein said second core recognition sequence is complementary to said first core recognition sequence.

1 more element

A composition for modulating transcription activation of trans-acting factor responsive genes, said composition comprising a double-stranded DNA fragment, wherein said DNA fragment contains at least one transcription control recognition sequence of at least 6 nucleotide base pairs, and wherein one strand of said DNA fragment is attached to the other strand by means of at least one linker covalently bound to each strand wherein said linker is selected from:

a sequence of bases sufficient to allow the formation of a loop structure between the 3'-end of one strand of said double-stranded DNA fragment and the 5'-end of the strand complementary thereto, or
a sequence of bases sufficient to allow the formation of a loop structure between the 5'-end of one strand of said double-stranded DNA fragment and the 3'-end of the strand complementary thereto, or
two sequences of bases sufficient to allow the formation of a loop structure between:
the 3'-end of the first strand of said double-stranded DNA fragment and the 5'-end of the strand complementary thereto and the 3'-end of the complementary strand of said double-stranded DNA fragment, and the 5'-end of the first strand of said double-stranded DNA fragment.

1 more element

Dependencies are inferred from the claim text.

Oligonucleotide decoys and methods relating thereto

Life of this patent

Oligonucleotide decoys and methods relating thereto

Life of this patent

Abstract

Claims20