AAV delivery of nucleobase editors

US11306324B2 · kind B2 · utility

Assignee

• President and Fellows of Harvard College · US

Inventors

• David R. Liu · Lexington, US
• Jonathan Ma Levy · Boston, US
• Wei Hsi Yeh · Cambridge, US

Key dates

• Filing date: Oct 13, 2017
• Grant date: Apr 19, 2022
• Expiry date: Aug 20, 2038

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2521/539
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

Provided herein are methods of delivering “split” Cas9 protein or nucleobase editors into a cell, e.g., via a recombinant adeno-associated virus (rAAV), to form a complete and functional Cas9 protein or nucleobase editor. The Cas9 protein or the nucleobase editor is split into two sections, each fused with one part of an intein system (e.g., intein-N and intein-C encoded by dnaEn and dnaEc, respectively). Upon co-expression, the two sections of the Cas9 protein or nucleobase editor are ligated together via intein-mediated protein splicing. Recombinant AAV vectors and particles for the delivery of the split Cas9 protein or nucleobase editor, and methods of using such AAV vectors and particles are also provided.