Altering microbial populations and modifying microbiota

Patent metadata

Field	Value
Publication number	US-12514869-B2
Application number	US-202519030161-A
Country	US
Kind code	B2
Filing date	Jan 17, 2025
Priority date	May 6, 2015
Publication date	Jan 6, 2026
Grant date	Jan 6, 2026

Abstract

The invention relates to methods, uses, systems, arrays, engineered nucleotide sequences and vectors for inhibiting bacterial population growth or for altering the relative ratio of sub-populations of first and second bacteria in a mixed population of bacteria. The invention is particularly useful, for example, for treatment of microbes such as for environmental, medical, food and beverage use. The invention relates inter alia to methods of controlling microbiologically influenced corrosion (MIC) or biofouling of a substrate or fluid in an industrial or domestic system.

First claim

What is claimed is:

1 . A method for modifying a mixed population of bacteria, wherein the mixed population of bacteria comprises a first bacterial sub-population and a second bacterial sub-population, wherein the first bacterial sub-population comprises a first bacterial species and the second bacterial sub-population comprises a population of host cells of a second bacterial species, wherein the second bacterial species is a different species than the first bacterial species, the method comprising:
producing a crRNA from an engineered nucleic acid in the host cells, wherein the crRNA hybridizes to a target sequence in the host cells and guides a Cas to modify the target sequence in the host cells, wherein the first bacterial species is a gram positive species and the second bacterial species is a gram positive species, wherein the growth of the first bacterial species in the mixed population is not inhibited, and wherein the host cells are killed or growth of the host cells is reduced.

2 . The method of claim 1 , wherein the crRNA is under control of an inducible promoter.

3 . The method of claim 1 , wherein the Cas is a Type I Cas.

4 . The method of claim 1 , wherein the Cas is a Type II Cas.

5 . The method of claim 1 , wherein the Cas is a Type III Cas.

6 . The method of claim 1 , wherein the Cas is encoded by an engineered nucleic acid.

7 . The method of claim 1 , wherein the Cas is a functional endogenous Cas of the host cells.

8 . The method of claim 1 , wherein the engineered nucleic acid for producing the crRNA is present in a phage, phagemid or plasmid.

9 . The method of claim 6 , wherein the engineered nucleic acid encoding the Cas is present in a phage, phagemid or plasmid.

10 . The method of claim 1 , wherein the mixed population of bacteria is present in a human microbiota.

11 . The method of claim 1 , wherein the mixed population of bacteria is present in a subject, and wherein the host cells cause a disease in the subject.

12 . The method of claim 1 , wherein the mixed population of bacteria is present in a subject, and wherein the host cells produce an exogenous protein.

13 . The method of claim 12 , wherein the exogenous protein is an antibiotic.

14 . The method of claim 1 , wherein the Cas is a Cas nuclease, and wherein the crRNA guides the Cas to cut the target sequence in the host cells.

15 . The method of claim 1 , wherein the mixed population of bacteria is a gut microbiota of a human or an animal.

16 . The method of claim 1 , wherein the mixed population of bacteria is present in an industrial or medical fluid, an apparatus, a container, a waterway, water, a beverage, a foodstuff, or a cosmetic, and wherein the host cells cause contamination of the industrial or medical fluid, apparatus, container, waterway, water, beverage, foodstuff or cosmetic.

17 . The method of claim 1 , wherein the mixed population of bacteria comprises a third bacterial species, wherein the third bacterial species is a human gut commensal species or a human gut probiotic species.

18 . The method of claim 1 , wherein the first bacterial species is L. lactis.

19 . The method of claim 15 , wherein the second bacterial species is Streptococcus.

20 . The method of claim 15 , wherein the mixed population of bacteria comprises a third bacterial species, wherein the third bacterial species is E. coli.

21 . The method of claim 1 , wherein the first bacterial species has a 16s ribosomal RNA-encoding DNA sequence that is at least about 80% identical to a 16s ribosomal RNA-encoding DNA sequence of the second bacterial species.

22 . A method for modifying a mixed population of bacteria, wherein the mixed population of bacteria comprises a first bacterial sub-population and a second bacterial sub-population, wherein the first bacterial sub-population comprises a first bacterial species and the second bacterial sub-population comprises a population of host cells of a second bacterial species, wherein the second bacterial species is a different species than the first bacterial species, the method comprising:
producing a crRNA from an engineered nucleic acid in the host cells, wherein the crRNA hybridizes to a target sequence in the host cells and guides a Cas to modify the target sequence in the host cells, wherein the first bacterial species has a 16s ribosomal RNA-encoding DNA sequence that is at least about 80% identical to a 16s ribosomal RNA-encoding DNA sequence of the second bacterial species, wherein the growth of the first bacterial species in the mixed population is not inhibited, and wherein the host cells are killed or growth of the host cells is reduced.

23 . A method for modifying a mixed population of bacteria, wherein the mixed population of bacteria comprises a first bacterial sub-population and a second bacterial sub-population, wherein the first bacterial sub-population comprises a first bacterial species and the second bacterial sub-population comprises a population of host cells of a second bacterial species, wherein the second bacterial species is a different species than the first bacterial species, the method comprising:
producing a crRNA from an engineered nucleic acid in the host cells, wherein the crRNA hybridizes to a target sequence in the host cells and guides a Cas in the host cells to modify the target sequence in the host cells, wherein the first bacterial species is a Firmicutes and the second bacterial species is a Firmicutes, wherein the growth of the first bacterial species in the mixed population is not inhibited, and wherein the host cells are killed or growth of the host cells is reduced.

Assignees

• Snipr Tech Ltd

Inventors

• Clube Jasper
• Sommer Morten
• Grøndahl Christian
• Van Der Helm Eric
• Vazquez-Uribe Ruben

Classifications

• A61K2300/00

  Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00 · CPC title

• C12N2795/00032

  Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• A61K48/005

  characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered · CPC title