Method for generating higher order genome editing libraries

US12091656B2 · kind B2 · utility

Assignee

• Johann Wolfgang Goethe-Universität Frankfurt am Main · DE

Inventors

• Manuel Kaulich · Frankfurt am Main, DE
• Andreas Ernst · Worms, DE
• Martin Wegner · Meckenheim, DE
• Valentina DIEHL · Frankfurt am Main, DE
• Rahel DE BRUYN · Heidingsfeld, DE
• Svenja WIECHMANN · Frankfurt am Main, DE

Key dates

• Filing date: Dec 27, 2017
• Grant date: Sep 17, 2024
• Expiry date: Dec 23, 2039

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Y605/01001
• WIPO field: Organic fine chemistry
• WIPO sector: Chemistry

Abstract

The present invention pertains to a novel method for the generation of highly diverse RNA expressing vectors and vector libraries for use in targeted gene knock out, knock down and genome modification approaches. The invention pertains to a method for generating such higher order libraries without the need of classical cloning technologies. This is particularly useful for libraries based on large vectors wherein a sequence cannot be easily mutated with classical mutagenesis methods. The vectors and libraries generated according to the methods of the invention are in particular for RNA assisted silencing technologies such as RNA interference, and for targeted genome editing using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system or similar RNA/DNA-encoded gene perturbation systems which use small guide RNAs to target the CRISPR complex to a specific genomic sequence. The invention provides also kits comprising the materials for performing the methods of the invention.