Methods for labeling a population of RNA molecules

US11479766B2 · kind B2 · utility

Assignee

• New England Biolabs, Inc. · US

Inventors

• Ira Schildkraut · Santa Fe, US
• Laurence Ettwiller · Ipswich, US
• Ivan R. Correa, Jr. · Ipswich, US
• George Tzertzinis · Cambridge, US
• John Buswell · Byfield, US
• Madalee G. Wulf · Ipswich, US

Key dates

• Filing date: Feb 28, 2018
• Grant date: Oct 25, 2022
• Expiry date: Aug 19, 2039

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6806
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A method of labeling, and optionally enriching, for a population of target RNA molecules in a mixture of RNAs is provided. In some embodiments, the method may comprise (a) adding a label to the 5′ end of 5′-diphosphorylated or 5′-triphosphorylated target RNA molecules in a sample by incubating the sample with labeled GTP and a capping enzyme; and (b) optionally enriching for target RNA comprising the affinity tag-labeled GMP using an affinity matrix that binds to the affinity tag. The label may be an oligonucleotide, which may further comprise an affinity group attached either internally or at 5′ or 3′ end of the oligonucleotide where the oligonucleotide label may be added directly, or indirectly via a reaction with a reactive group to the target RNA.