Methods and compositions for killing a target bacterium
AssigneesLocus Biosciences, Inc., North Carolina State University
InventorsPaul M. GAROFOLO, David G. OUSTEROUT, Kurt SELLE, Sandi WONG, Chase Lawrence BEISEL, Atul Kumar SINGH
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Abstract
Provided herein are methods and compositions for killing a target bacterium with a CRISPR-Cpf1 system. Also disclosed are engineered bacteriophages.
Claims216
A method for killing a target bacterium comprising:
- introducing into a target bacterium a bacteriophage comprising:
- (a) a first nucleic acid encoding a spacer sequence or a crRNA transcribed therefrom, wherein the spacer sequence is complimentary to a target nucleotide sequence from a target gene in the target bacterium;
- and
- (b) a second nucleic acid encoding a transcriptional activator for a CRISPR-Cpf1 system in a target bacterium;
- wherein the target bacterium is killed by lytic activity of the bacteriophage or activity of a CRISPR-Cpf1 system using the spacer sequence or the crRNA transcribed therefrom.
The method of any one claims of 1-19, wherein the target bacterium is killed solely by the activity of the CRISPR-Cpf1 system.
A bacteriophage comprising:
- (a) a first nucleic acid encoding a spacer sequence or a crRNA transcribed therefrom, wherein the spacer sequence is complimentary to a target nucleotide sequence from a target gene in a target bacterium;
- and
- (b) a second nucleic acid encoding a encoding a transcriptional activator for a CRISPR-Cpf1 system in a target bacterium,
- wherein the target bacterium is killed by the lytic activity of the bacteriophage or activity of a CRISPR-Cpf1 system using the spacer sequence or the crRNA transcribed therefrom.
A method for modulating the activity of a CRISPR-Cpf1 system in a bacterium, comprising:
- introducing a bacteriophage comprising a nucleic acid encoding a transcriptional activator for the CRISPR-Cpf1 system in the target bacterium.
A bacteriophage comprising a nucleic acid encoding a transcriptional activator for a CRISPR-Cpf1 system in a target bacterium.
A method of killing a target bacterium, comprising introducing into a target bacterium a bacteriophage comprising:
- (a) lytic activity, and
- (b) a first nucleic acid sequence encoding an anti-CRISPR polypeptide,
- wherein the anti-CRISPR polypeptide enhances the lytic activity of the bacteriophage.
A bacteriophage comprising:
- (a) lytic activity, and
- (b) a first nucleic acid sequence encoding an anti-CRISPR polypeptide,
- wherein the anti-CRISPR polypeptide enhances the lytic activity of the bacteriophage.
A method for killing a target bacterium comprising:
- introducing into a target bacterium a bacteriophage comprising:
- (a) a first nucleic acid encoding a spacer sequence or a crRNA transcribed therefrom, wherein the spacer sequence is complimentary to a target nucleotide sequence from a target gene in the target bacterium;
- and
- (b) a second nucleic acid encoding an exogenous Cpf1;
- wherein the target bacterium is killed by lytic activity of the bacteriophage or activity of a CRISPR-Cpf1 system using the spacer sequence or the crRNA transcribed therefrom and the exogenous Cpf1.
The method of any one claims of 139-147, wherein the target bacterium is killed solely by the activity of the CRISPR-Cpf1 system.
A bacteriophage comprising:
- (a) a first nucleic acid encoding a spacer sequence or a crRNA transcribed therefrom, wherein the spacer sequence is complimentary to a target nucleotide sequence from a target gene in a target bacterium;
- and
- (b) a second nucleic acid encoding a encoding an exogenous Cpf1,
- wherein the target bacterium is killed by the lytic activity of the bacteriophage or activity of a CRISPR-Cpf1 system using the spacer sequence or the crRNA transcribed therefrom and the exogenous Cpf1.
A method for modulating the activity of a CRISPR-Cpf1 system in a bacterium, comprising:
- introducing a bacteriophage comprising a nucleic acid encoding an exogenous Cpf1 for the CRISPR-Cpf1 system in the target bacterium.
A bacteriophage comprising a nucleic acid encoding an exogenous Cpf1 for a CRISPR-Cpf1 system in a target bacterium.
Dependencies are inferred from the claim text.