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.
Claims18
A pharmaceutical composition for killing targeted bacteria in a mixed bacterial population comprising:
- a pharmaceutically acceptable carrier and a clustered regularly interspaced short palindromic repeats (CRISPR) system packaged in a phage,
- wherein the CRISPR system comprises DNA encoding:
- i) a Type I, Type II, or Type III 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 CRISPR system is introduced into bacteria in the bacterial population,
- wherein subsequent to the introduction of the CRISPR system, the targeting RNA and the CRISPR-associated enzyme are expressed in the bacteria into which the CRISPR system is introduced,
- wherein the expressed 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 clustered regularly interspaced short palindromic repeats (CRISPR) system packaged in a phage,
- wherein the CRISPR system comprises DNA encoding:
- i) a Type I, Type II, or Type III 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 CRISPR system is introduced into bacteria in the bacterial population,
- wherein subsequent to the introduction of the CRISPR system, the targeting RNA and the CRISPR-associated enzyme are expressed in the bacteria into which the CRISPR system is introduced,
- wherein the expressed 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.
Dependencies are inferred from the claim text.