Sequence specific antimicrobials
AssigneeThe Rockefeller University
InventorsDavid Bikard, Luciano Marraffini
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Abstract
Provided are compositions and methods for selectively reducing the amount of antibiotic resistant and/or virulent bacteria in a mixed bacteria population, or for reducing any other type of unwanted bacteria in a mixed bacteria population. The compositions and methods involve targeting bacteria that are differentiated from other members of the population by at least one unique clustered regularly interspaced short palindromic repeats (CRISPR) targeted DNA sequence. The compositions and methods can be readily adapted to target any bacteria or any bacteria plasmid, or both.
Claims25
A pharmaceutical composition for killing targeted bacteria in a mixed bacterial population comprising:
- a pharmaceutically acceptable carrier and
- a recombinant phagemid,
- wherein the recombinant phagemid comprises a clustered regularly interspaced short palindromic repeats (CRISPR) system,
- wherein the CRISPR system comprises DNA encoding:
- i) a Type II CRISPR-associated enzyme;
- and ii) a targeting RNA that targets at least one bacterial chromosome at a target site;
- and
- wherein, upon contacting a bacterial population containing the at least one bacterial chromosome with the pharmaceutical composition, the recombinant phagemid is introduced into bacteria in the bacterial population,
- wherein subsequent to the introduction of the recombinant phagemid, the targeting RNA and the Type II CRISPR-associated enzyme are expressed in the bacteria into which the recombinant phagemid is introduced,
- wherein the expressed Type II CRISPR-associated enzyme cleaves the bacterial chromosome at the target site of the targeting RNA, and
- wherein the cleavage of the bacterial chromosome at the target site kills the bacteria.
A pharmaceutical composition for killing targeted bacteria in a mixed bacterial population comprising:
- a pharmaceutically acceptable carrier and
- a recombinant phagemid,
- wherein the recombinant phagemid comprises a clustered regularly interspaced short palindromic repeats (CRISPR) system,
- wherein the CRISPR system comprises DNA encoding:
- i) a Type II CRISPR-associated enzyme;
- and ii) a targeting RNA that targets an antibiotic resistance gene on a bacterial plasmid at a target site within the bacterial plasmid;
- wherein, upon contacting a bacterial population containing the antibiotic resistance gene on the bacterial plasmid with the pharmaceutical composition, the recombinant phagemid is introduced into bacteria in the bacterial population,
- wherein the targeting RNA and the Type II CRISPR-associated enzyme are expressed in the bacteria into which the recombinant phagemid is introduced,
- wherein the expressed Type II CRISPR-associated enzyme cleaves the antibiotic resistance gene on a bacterial plasmid at the target site within the bacterial plasmid, and
- wherein the cleavage of the bacterial plasmid at the target site kills the bacteria in the presence of the antibiotic.
A pharmaceutical composition for killing targeted bacteria in a mixed bacterial population comprising:
- a pharmaceutically acceptable carrier and
- a recombinant phagemid,
- wherein the recombinant phagemid comprises a clustered regularly interspaced short palindromic repeats (CRISPR) system,
- wherein the CRISPR system comprises DNA encoding:
- i) a Type II CRISPR-associated enzyme;
- and ii) a targeting RNA that targets a bacterial plasmid comprising an antibiotic resistance gene at a target site within the bacterial plasmid;
- wherein, upon contacting a bacterial population containing the bacterial plasmid comprising an antibiotic resistance gene with the pharmaceutical composition, the recombinant phagemid is introduced into bacteria in the bacterial population,
- wherein the targeting RNA and the Type II CRISPR-associated enzyme are expressed in the bacteria into which the recombinant phagemid is introduced,
- wherein the expressed Type II CRISPR-associated enzyme cleaves the bacterial plasmid comprising an antibiotic resistance gene at the target site within the bacterial plasmid, and
- wherein the cleavage of the bacterial plasmid at the target site kills the bacteria in the presence of the antibiotic.
Dependencies are inferred from the claim text.