Modified triple-helix forming oligonucleotides for targeted mutagenesis

US8658608B2 · kind B2 · utility

Assignees

• Yale University · US
• The United States of America, as represented by the Department of Health and Human Services · US

Inventors

• Peter Glazer · Guilford, US
• Michael M. Siedman · Washington, US

Key dates

• Filing date: Nov 22, 2006
• Grant date: Feb 25, 2014
• Expiry date: Mar 25, 2028

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2320/13
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

High affinity, chemically modified triplex-forming oligonucleotides (TFOs) and methods for use thereof are disclosed. TFOs are defined as triplex-forming oligonucleotides which bind as third strands to duplex DNA in a sequence specific manner. Triplex-forming oligonucleotides may be comprised of any possible combination of nucleotides and modified nucleotides. Modified nucleotides may contain chemical modifications of the heterocyclic base, sugar moiety or phosphate moiety. A high affinity oligonucleotide (Kd≦2×10−8) which forms a triple strand with a specific DNA segment of a target gene DNA is generated. It is preferable that the Kd for the high affinity oligonucleotide is below 2×10−10. The nucleotide binds or hybridizes to a target sequence within a target gene or target region of a chromosome, forming a triplex region. The binding of the oligonucleotide to the target region stimulates mutations within or adjacent to the target region using cellular DNA synthesis, recombination, and repair mechanisms. The mutation generated activates, inactivates, or alters the activity and function of the target gene.