BACTERIOPHAGES FOR REDUCING TOXICITY OF BACTERIA

• US 20130315869A1
• Filed: 05/22/2012
• Published: 11/28/2013
• Est. Priority Date: 05/22/2012
• Status: Active Grant

• Current Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Original Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Inventors: Friedman, Nir, QIMRON, Ehud, Edgar, Rotem
• Granted Patent: US 8,865,158 B2
• Time in Patent Office: Days
• US Class Current: 424/93.2
• CPC Class Codes: A61K 35/76 Viruses; Subviral particles; Bacteriophages
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: C12N 2795/10332 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent

Abstract

A genetically modified bacteriophage is disclosed which comprises:

• • (i) an exogenous polynucleotide which encodes an agent which reduces the toxicity of a bacterium; and
  • (ii) an exogenous polynucleotide which encodes a selectable marker.

Uses thereof and kits comprising same are also disclosed.

Claims

1. A genetically modified bacteriophage comprising:

(i) an exogenous polynucleotide which encodes an agent which reduces the toxicity of a bacterium, wherein said agent is non-toxic to the bacterium and does not affect directly the survival of said bacterium; and

(ii) an exogenous polynucleotide which encodes a selectable marker that renders the bacterium infected by the bacteriophage insensitive to a compound which is toxic to bacteria, wherein said selectable marker is not an antibiotic resistance gene.

2. A kit comprising a compound which is toxic to bacteria and the phage of claim 1, wherein said selectable marker renders a bacterium infected by said phage insensitive to said compound.

4. The bacteriophage of claim 1, wherein said agent reverses resistance of said bacterium to an antibiotic.

5. The bacteriophage of claim 4, wherein said resistance is due to a mutated polypeptide of said bacterium selected to be 30S ribosomal subunit protein S12.

6. The bacteriophage of claim 1, wherein said agent is non-toxic to the bacterium.

7. The bacteriophage of claim 4, wherein said agent comprises a polypeptide selected to be 30S ribosomal subunit protein S12.

8. The bacteriophage of claim 7, wherein said agent is a 30S ribosomal subunit protein S12.

9. The bacteriophage of claim 4, wherein said agent that reverses resistance of said bacterium to an antibiotic is 30S ribosomal subunit protein S12, encoded by the polynucleotide sequence of SEQ ID NO:

24.

10. The bacteriophage of claim 1, wherein said agent is a polynucleotide agent which down-regulates expression of an antibiotic resistance gene expressed in said bacterium.

11. The bacteriophage of claim 1, wherein said agent is a polynucleotide agent which down-regulates expression of a virulence gene expressed in said bacterium.

12. The bacteriophage of claim 10, wherein said polynucleotide agent is selected from the group consisting of an siRNA, a short hairpin RNA, a ribozyme and a DNAzyme.

18. The bacteriophage of claim 1, wherein said selectable marker comprises a resistance marker to tellurite.

20. The bacteriophage of claim 1, being a lambda temperate phage.

22. An anti-bacterial composition, comprising a carrier and as an active ingredient the bacteriophage of claim 1.

24. The antibacterial composition of claim 22, formulated as a spray, a stick, a paint, a gel, a cream, wash, a wipe, a foam, a soap, an oil, a solution, a lotion, an ointment or a paste.

26. A method of preventing a bacterial infection which is resistant to an antibiotic in a subject, the method comprising contacting a solid surface with an anti-bacterial composition that includes the bacteriophage of claim 1, thereby preventing the bacterial infection.

28. The method of claim 26, further comprising contacting the solid surface with a compound which is toxic to bacteria.

31. An isolated population of bacterial cells comprising the bacteriophage of claim 1.

3. (canceled)

13. A genetically modified bacteriophage comprising an exogenous polynucleotide which encodes a polypeptide which reverses resistance of a bacterium to an antibiotic, wherein said polypeptide is 30S ribosomal subunit protein S12.

14. The bacteriophage of claim 13, wherein said polypeptide is non-toxic to the bacterium.

15. The bacteriophage of claim 13, wherein said polypeptide comprises a 30S ribosomal subunit protein S12.

16. The bacteriophage of claim 13, wherein said 30S ribosomal subunit protein S12 is encoded by the polynucleotide sequence of SEQ ID NO:

24.

17. The bacteriophage of claim 13, wherein said exogenous polynucleotide further encodes a selectable marker.

19. The bacteriophage of claim 17, wherein said selectable marker comprises a resistance marker to tellurite.

21. The bacteriophage of claim 13, being a lambda temperate phage.

23. An anti-bacterial composition, comprising a carrier and as an active ingredient the bacteriophage of claim 13.

25. The antibacterial composition of claim 23, formulated as a spray, a stick, a paint, a gel, a cream, wash, a wipe, a foam, a soap, an oil, a solution, a lotion, an ointment or a paste.

27. A method of preventing a bacterial infection which is resistant to an antibiotic in a subject, the method comprising contacting a solid surface with an anti-bacterial composition that includes the bacteriophage of claim 13, thereby preventing the bacterial infection.

29. The method of claim 27, further comprising contacting the solid surface with a compound which is toxic to bacteria.

30. The method of claim 29, wherein said exogenous polynucleotide further encodes a selectable marker that renders a bacterium infected by said phage insensitive to said compound.