Methods and compositions for using argonaute to modify a single stranded target nucleic acid

US10253311B2 · kind B2 · utility

Assignee

• The Regents of the University of California · US

Inventors

• Jennifer A. Doudna · Berkeley, US
• Emine Kaya · Oakland, US
• Kilian R. Knoll · Berkeley, US

Key dates

• Filing date: Apr 9, 2015
• Grant date: Apr 9, 2019
• Expiry date: Apr 9, 2035

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12P19/34
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present disclosure provides compositions, kits, genetically modified cells, non-human transgenic organisms, and methods for binding and/or cleaving a single stranded target nucleic acid. A method of cleaving includes contacting a single stranded target nucleic acid with (e.g., introducing into a cell) a subject argonaute (Ago) polypeptide and a guide RNA (e.g., having a 5′-OH). In some embodiments, a subject Ago polypeptide includes an amino acid sequence having 70% or more sequence identity with amino acids 282-430 and/or 431-639 of the Marinitoga piezophila argonaute (MpAgo) protein set forth in SEQ ID NO: 1. The present disclosure provides variant Ago polypeptides; and methods of use of same.