Granted Patent B2

US 12,295,993 · App. 17/088,302 · Granted May 13, 2025

Inventors: David Bikard (Paris, FR); Luciano Marraffini (Brooklyn, NY)

Assignee: The Rockefeller University

A61K38/465A01N63/00A61K31/7105A61K31/713A61K45/06C12N9/16C12N9/22C12N15/113C12N15/74C12Y301/00C12N2310/10C12N2310/20C12N2795/10331C12N2795/10332C12N2795/10343C12N2795/10371

Abstract

Claims (16)

1. A method for killing targeted pathogenic bacteria in a mixed bacterial population in a subject comprising:

providing a pharmaceutical composition comprising a pharmaceutically acceptable carrier and packaged, recombinant phagemids that are packaged in phage capsids,

wherein the packaged phagemids comprise 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

contacting the bacterial population with the pharmaceutical composition by oral, parenteral, topical, nasal, or rectal administration to the subject;

wherein the contacting with the pharmaceutical composition introduces at least some of the phagemids into at least some of the bacteria in the bacterial population,

wherein subsequent to the introduction of the phagemids, the bacteria into which the phagemid is introduced expresses the targeting RNA and the type II CRISPR-associated enzyme,

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 targeted pathogenic bacteria.

2. The method of claim 1 , wherein the CRISPR-associated enzyme is a Cas9 enzyme.

3. The method of claim 2 , wherein the Cas9 enzyme is a Streptococcus pyogenes Cas9.

4. The method of claim 1 , further comprising detecting the killing of bacteria by the pharmaceutical composition.

5. The method of claim 1 , wherein the mixed bacterial population comprises a combination of bacterial species selected from the group consisting of Staphylococcus, Clostridium, Bacillus, Salmonella, Helicobacter pylori, Neisseria gonorrhoeae, Neisseria meningitidis , and Escherichia coli.

6. The method of claim 1 , wherein the targeted bacteria is a Staphylococcus.

7. The method of claim 1 , wherein the targeted bacteria is a Mycobacterium.

8. The method of claim 1 , wherein the targeted bacteria is a Pseudomonas.

Assignments (2)

Continuity (8)

References Cited (55)