The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Oct. 10, 2017

Filed:

Aug. 26, 2015
Applicant:

Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt, Neuherberg, DE;

Inventors:

Ralf Kühn, Freising, DE;

Wolfgang Wurst, München, DE;

Oskar Ortiz Sanchez, München, DE;

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
A01N 63/00 (2006.01); A61K 48/00 (2006.01); C12N 5/075 (2010.01); A01K 67/027 (2006.01); C12N 9/22 (2006.01); C12N 15/89 (2006.01); C12N 15/90 (2006.01); A61D 19/04 (2006.01); C12N 15/10 (2006.01);
U.S. Cl.
CPC ...
C12N 5/0609 (2013.01); A01K 67/0275 (2013.01); A61D 19/04 (2013.01); C12N 9/22 (2013.01); C12N 15/102 (2013.01); C12N 15/89 (2013.01); C12N 15/907 (2013.01); A01K 2217/072 (2013.01); A01K 2217/075 (2013.01); A01K 2227/105 (2013.01); A01K 2267/03 (2013.01); C12N 2800/80 (2013.01);
Abstract

The present invention relates to a method of producing a non-human, mammalian oocyte carrying a modified target sequence in its genome, the method comprising the steps of introducing into a non-human, mammalian oocyte: (a) a clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated protein 9 (Cas9 protein) or a nucleic acid molecule encoding said Cas9 protein; and (b-i) a target sequence specific CRISPR RNA (crRNA) and a trans-activating crRNA (tracr RNA) or a nucleic acid molecule encoding said RNAs; or (b-ii) a chimaeric RNA sequence comprising a target sequence specific crRNA and tracrRNA or a nucleic acid molecule encoding said RNA; wherein the Cas9 protein introduced in (a) and the RNA sequence(s) introduced in (b-i) or (b-ii) form a protein/RNA complex that specifically binds to the target sequence and introduces a single or double strand break within the target sequence. The present invention further relates to the method of the invention, wherein the target sequence is modified by homologous recombination with a donor nucleic acid sequence further comprising the step: (c) introducing a nucleic acid molecule into the cell, wherein the nucleic acid molecule comprises the donor nucleic acid sequence and regions homologous to the target sequence. The present invention also relates to a method of producing a non-human mammal carrying a modified target sequence in its genome.


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