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1. WO1996040898 - TRIPLE-HELIX FORMING OLIGONUCLEOTIDES FOR TARGETED MUTAGENESIS

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[ EN ]
We claim:
1. A pharmaceutical composition comprising a high affinity oligonucleotide for targeted mutagenesis of a double-stranded nucleic acid molecule, the single-stranded oligonucleotide having a sequence that forms a triple-stranded nucleic acid molecule with a target region of the double-stranded nucleic acid molecule, wherein the oligonucleotide has a K,, of 2 x 10 "8 or less for the target region.
2. The composition of claim 1 wherein the oligonucleotide is at least 20 nucleotide residues in length.
3. The composition of claim 1 further comprising a triple-helix forming oligonucleotide linked to a mutagenic compound.
4. The composition of claim 1 wherein the double-stranded nucleic acid molecule encodes a molecule and the targeted mutagenesis alters the activity of the molecule encoded by the double-stranded nucleic acid molecule.
5. The composition of claim 4 wherein the double-stranded nucleic acid molecule is a gene.
6. The composition of claim 5 wherein the gene is an oncogene.

7. The composition of claim 5 wherein the gene is a defective gene.
8. The composition of claim 4 wherein the double-stranded nucleic acid molecule is all or a portion of a viral genome.
9. The composition of claim 1 further comprising a recombination fragment.
10. A method of producing a high affinity, triple-helix forming oligonucleotide comprising the step of synthesizing an oligonucleotide substantially complementary, based on the third strand binding code, to a target region of a double-stranded nucleic acid molecule, wherein the disassociation constant for the oligonucleotide and the target region is less than or equal to 2 x 10"8.
11. The method of claim 10, wherein the oligonucleotide is at least 20 nucleotide residues in length.

12. A method for targeted mutagenesis of a nucleic acid molecule comprising the steps of:
a) hybridizing a high affinity, triple-helix forming oligonucleotide to a target region of a double-stranded nucleic acid molecule, wherein the oligonucleotide comprises a single-stranded nucleic acid that forms a triple-stranded nucleic acid molecule with the target region; and
b) mutating the double-stranded nucleic acid molecule, wherein the disassociation constant for the oligonucleotide and the target region is less than or equal to 2 x 108.
13. The method of claim 12, wherein the mutagenic
oligonucleotide is at least 20 nucleotide residues in length.
14. The method of claim 12 wherein the double-stranded molecule encodes a molecule and the mutation alters the activity of the molecule encoded by the double-stranded nucleic acid molecule.
15. The method of claim 13 wherein the double-stranded nucleic acid molecule is a gene.
16. The method of claim 15 wherein the gene is an oncogene.
17. The method of claim 15 wherein the gene is a defective gene.

18. The method of claim 17 wherein the gene is a defective human β-hemoglobin gene.
19. The method of claim 14 wherein the double-stranded nucleic acid molecule is all or a portion of a viral genome.