Method for the site-specific enzymatic labelling of nucleic acids in vitro by incorporation of unnatural nucleotides

US10626138B2 · kind B2 · utility

Assignees

• THE SCRIPPS RESEARCH INSTITUTE NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT OF HEALTH AND HUMAN SERVICES (DHHS) · US
• U.S. GOVERNMENT NIH DIVISION OF EXTRAMURAL INVENTIONS AND TECHNOLOGY RESOURCES (DEITR) · US

Inventors

• Floyd E. ROMESBERG · Village of La Jolla, US
• Denis Malyshev · Solana Beach, US
• Lingjun Li · San Jose, US
• Thomas Lavergne · Le Versoud, FR
• Zhengtao LI · Shanghai, CN

Key dates

• Filing date: Aug 8, 2014
• Grant date: Apr 21, 2020
• Expiry date: May 27, 2036

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6832
• WIPO field: Organic fine chemistry
• WIPO sector: Chemistry

Abstract

Provided herein are analogs of unnatural nucleotides bearing predominantly hydrophobic nucleobase analogs that form unnatural base pairs during DNA polymerase-mediated replication of DNA or RNA polymerase-mediated transcription of RNA. In this manner, the unnatural nucleobases can be introduced in a site-specific way into oligonucleotides (single or double stranded DNA or RNA), where they can provide for site-specific cleavage, or can provide a reactive linker than can undergo functionalization with a cargo-bearing reagent by means of reaction with a primary amino group or by means of click chemistry with an alkyne group of the unnatural nucleobase linker.