Direct, programmable detection of epigenetic DNA cytosine modifications using TAL effectors

US9879315B2 · kind B2 · utility

Assignee

• University of Konstanz · DE

Inventors

• Daniel Summerer · Konstanz, DE
• M. Grzegorz Kubik · Konstanz, DE
• Moritz Schmidt · Cologne, DE

Key dates

• Filing date: Jun 26, 2014
• Grant date: Jan 30, 2018
• Expiry date: Jun 26, 2034

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6858
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Methods for the differentiation of a 5-hydroxymethyl modification of a cytosine residue of interest in a nucleic acid molecule from (i) a 5-methyl modification of the cytosine residue of interest or (ii) an unmodified cytosine residue of interest, the methods making use of the capability of transcription-activator-like effector (TALE) proteins to preferentially bind with strong affinity to nucleic acid sequences containing non-modified cytosine residues or 5-methyl modified cytosine residues, and to bind, if at all, with only strongly reduced affinity to nucleic acid sequences containing 5-hydroxymethyl modified cytosine residues. The present invention further relates to respective uses of TALE proteins for the differentiation of a 5-hydroxymethyl modification of a cytosine residue of interest in a nucleic acid molecule from (i) a 5-methyl modification of the cytosine residue of interest or (ii) the unmodified cytosine residue of interest.