US 12,285,467 B2
Sequence specific antimicrobials
David Bikard, Paris (FR); and Luciano Marraffini, Brooklyn, NY (US)
Assigned to The Rockfeller University, New York, NY (US)
Filed by The Rockefeller University, New York, NY (US)
Filed on Nov. 3, 2020, as Appl. No. 17/088,297.
Application 15/159,929 is a division of application No. 14/766,675, granted, now 10,660,943, issued on May 26, 2020, previously published as PCT/US2014/015252, filed on Feb. 7, 2014.
Application 17/088,297 is a division of application No. 16/877,010, filed on May 18, 2020.
Application 16/877,010 is a division of application No. 16/877,030, filed on May 18, 2020, granted, now 11,135,273.
Application 17/088,297 is a continuation of application No. 15/159,929, filed on May 20, 2016, granted, now 11,918,631.
Claims priority of provisional application 61/761,971, filed on Feb. 7, 2013.
Prior Publication US 2021/0060140 A1, Mar. 4, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 38/46 (2006.01); A01N 63/00 (2020.01); A61K 31/7105 (2006.01); A61K 31/713 (2006.01); A61K 45/06 (2006.01); C12N 9/16 (2006.01); C12N 9/22 (2006.01); C12N 15/113 (2010.01); C12N 15/74 (2006.01)
CPC A61K 38/465 (2013.01) [A01N 63/00 (2013.01); A61K 31/7105 (2013.01); A61K 31/713 (2013.01); A61K 45/06 (2013.01); C12N 9/16 (2013.01); C12N 9/22 (2013.01); C12N 15/113 (2013.01); C12N 15/74 (2013.01); C12Y 301/00 (2013.01); C12N 2310/10 (2013.01); C12N 2310/20 (2017.05); C12N 2795/10331 (2013.01); C12N 2795/10332 (2013.01); C12N 2795/10343 (2013.01); C12N 2795/10371 (2013.01)] 14 Claims
 
1. A method for killing targeted bacteria in a mixed bacterial population 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,
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 bacteria;
wherein the mixed bacterial population consists of two different strains of Staphylococcus aureus.