Phage-derived particles for in situ delivery of DNA payload into C. acnes population

Patent metadata

Field	Value
Publication number	US-11820989-B2
Application number	US-202117518936-A
Country	US
Kind code	B2
Filing date	Nov 4, 2021
Priority date	Nov 4, 2020
Publication date	Nov 21, 2023
Grant date	Nov 21, 2023

Abstract

The invention relates to C. acnes carrying DNA vectors with a C. acnes phage packaging signal and a gene of interest. The invention encompasses a C. acnes producer cell carrying DNA vectors, with a C. acnes phage packaging signal and a gene of interest, for the production of phage-derived particles that can robustly transduce C. acnes receiver cell allowing transgene expression. The invention encompasses C. acnes phage-derived particles carrying these vectors, C. acnes containing these vectors or modified by transduction of these phage-derived particles, and methods of using these phage-derived particles.

First claim

We claim:

1. A phagemid lacking genes encoding phage structural proteins comprising:
a phage packaging signal allowing packaging of the phagemid in a Cutibacterium acnes phage capsid,
wherein the phage packaging signal is at least 87% identical to SEQ ID NO: 76, and
a gene of interest that is a transgene that is exogenous to C. acnes;
wherein said phagemid is packaged in a C. acnes phage capsid.

2. The phagemid of claim 1 further comprising an origin of replication for C. acnes and a selection marker for C. acnes.

3. The phagemid of claim 1 , wherein the phagemid also comprises a CRISPR-Cas system.

4. The phagemid of claim 1 , wherein the phagemid comprises a template for homologous recombination with C. acnes phages.

5. The phagemid DNA vector of claim 1 , wherein the phagemid comprises a template for homologous recombination with C. acnes plasmids.

6. The phagemid of claim 3 , wherein the phagemid comprises a template for homologous recombination and wherein the CRISPR-Cas system targets the phagemid itself.

7. The phagemid of claim 2 , wherein the selection marker is not ermE.

8. The phagemid of claim 2 , wherein the selection marker is catA.

9. The phagemid of claim 1 , which comprises a DNA encoding an antigen.

10. An engineered C. acnes comprising the phagemid of claim 1 .

11. The engineered C. acnes according to claim 10 , wherein the phagemid comprises a DNA encoding an antigen.

12. An engineered C. acnes produced by contacting C. acnes with the phagemid of claim 1 , modifying the C. acnes with a gene of interest carried by the phagemid, selecting for the modification, and curing the C. acnes of the phagemid.

13. The engineered C. acnes of claim 10 , wherein the C. acnes has been modified by a CRISPR-Cas system carried by the phagemid.

14. The engineered C. acnes of claim 10 , wherein the C. acnes has been modified by insertion of an exogenous gene into the C. acnes chromosome.

15. A method for engineering a C. acnes comprising introducing the phagemid of claim 1 into a C. acnes.

16. The method of claim 15 , further comprising selecting a modified C. acnes.

17. The method of claim 16 , comprising selecting a modified C. acnes that has an insertion of an exogenous gene into the C. acnes chromosome.

18. A vaccine and/or immunogenic composition comprising the engineered C. acnes of claim 10 comprising a phagemid comprising a nucleic acid encoding an antigen.

Assignees

• Eligo Bioscience

Inventors

• Leveau Aymeric
• Canadas Blasco Inès
• Mathieu Aurélie
• Decrulle Antoine

Classifications

• C12N15/76 Primary

  for Actinomyces; for Streptomyces · CPC title

• A61K39/02

  Bacterial antigens · CPC title

• A61K39/05

  Actinobacteria, e.g. Actinomyces, Streptomyces, Nocardia, Bifidobacterium, Gardnerella}, Corynebacterium; Propionibacterium {(Mycobacterium A61K39/04) · CPC title

• A61P17/10

  Anti-acne agents · CPC title

• C12N9/22

  Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title