Anti-CRISPR genes and proteins and methods of use
AssigneesDupont Nutrition Biosciences Aps, Universite Laval (Canada)
InventorsChristophe Fremaux, Philippe Horvath, Dennis A. Romero, Sylvain Moineau, Alexander Hynes, Geneviève Rousseau
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Abstract
The invention relates to anti-CRISPR genes and anti-CRISPR proteins, and their uses in various biotechnology applications.
Claims14
A method to favor the screening of a bacterial mechanism providing resistance against a virulent phage other than a given class 2 type II CRISPR-Cas mediated resistance, comprising:
- 1) providing a bacterial strain, the genome of which contains a given class 2 type II CRISPR-Cas system, which is known to be active against target nucleic acids;
- 2) expressing in said bacterial strain a gene encoding a protein which interferes with a function of said given class 2 type II CRISPR-Cas system;
- 3) exposing said bacterial strain of step 2) to a virulent phage;
- and
- 4) selecting bacteriophage-insensitive mutants (BIMs),
- wherein the resistance in said selected BIMs is provided by a bacterial mechanism other than the given class 2 type II CRISPR-Cas-mediated resistance.
A method to enrich a bacterial population in bacteriophage-insensitive mutants (BIMs) other than BIMs due to a given CRISPR-Cas system, comprising:
- 1) providing a bacterial population comprising a bacterial strain, the genome of which contains a given class 2 type II CRISPR-Cas system, which is known to be active against target nucleic acids;
- 2) expressing in said bacterial strain a gene encoding a protein which interferes with a function of said given class 2 type II CRISPR-Cas system;
- 3) exposing said bacterial strain of step 2) to a virulent phage;
- and
- 4) selecting BIMs,
- wherein the selected BIMs have acquired resistance to said phage by a mechanism other than the given class 2 type II CRISPR-Cas system.
A method to downmodulate, in a bacterial strain, the activity of a class 2 type II CRISPR-Cas system against target nucleic acids, comprising expressing in said bacterial strain a gene encoding a protein which interferes with a function of said class 2 type II CRISPR-Cas system, wherein said gene encodes a protein having at least 70% similarity or at least 90% identity to the sequence defined in SEQ ID NO:
- 10.
Dependencies are inferred from the claim text.