Altering microbial populations and modifying microbiota
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.
1. A method for modifying a target sequence in a bacterial or archaeal host cell comprising an endogenous nucleic acid functional to express an endogenous Type II Cas having nuclease activity in the host cell, the method comprising:
(a) contacting the host cell with an engineered nucleic acid for producing a host modifying crRNA (HM-crRNA), wherein the engineered nucleic acid does not encode the Type II Cas, and
(b) producing the HM-crRNA in the host cell;
wherein
(i) the HM-crRNA is operable with the endogenous Type II Cas expressed in the host cell, wherein the engineered nucleic acid and the Type II Cas are comprised by a Type II HM-CRISPR/Cas system in the host cell;
(ii) the HM-crRNA comprises a nucleotide sequence that is capable of hybridizing to the target sequence in the host cell to guide the endogenous Type II Cas to modify the target sequence in the host cell;
wherein the target sequence is modified by the Type II HM-CRISPR/Cas system; and
wherein the host cell is killed or growth of the host cell is inhibited.
2. The method of claim 1 , wherein the Type II Cas is a Cas9.
3. The method of claim 2 , wherein the Type II Cas is a Streptococcus Cas9.
4. The method of claim 1 , wherein the engineered nucleic acid for producing the HM-crRNA is present in a phage, phagemid or plasmid.
5. The method of claim 1 , wherein the target sequence is a host target sequence.
6. The method of claim 1 , wherein the Type II HM-CRISPR/Cas system comprises an endogenous tracrRNA of the host cell.
7. The method of claim 1 , wherein the Type II HM-CRISPR/Cas system comprises a tracrRNA, and wherein the tracrRNA is encoded by an engineered nucleic acid.
8. The method of claim 7 , wherein the engineered nucleic acid for producing the HM-crRNA encodes a single guide RNA comprising the tracrRNA and the HM-crRNA.
9. The method of claim 1 , wherein the host cell is a C. difficile, Klebsiella, Pseudomonas aeruginosa, Helicobacter pylori or Salmonella cell.
10. The method of claim 2 , wherein the Type II Cas is a spCas9 or saCas9.
11. The method of claim 1 , wherein the method is for medical, dental or ophthalmic use.
12. The method of claim 1 , wherein the host cell is comprised by a plant.
13. The method of claim 12 , wherein the plant is a crop.
14. The method of claim 1 , wherein the host cell is comprised by an animal.
15. The method of claim 14 , wherein the animal is a human.
16. The method of claim 1 , wherein the host cell is a cell type found in human microbiota.
17. The method of claim 1 , wherein the method is for medical, dental or ophthalmic use, and wherein the host cell is comprised by an animal or a human.
18. The method of claim 1 , wherein the method is for environmental or agricultural use, and wherein the host cell is comprised by a plant.
19. The method of claim 1 , wherein the host cell is a type of cell that infects an organism selected from a plant or an animal.
20. The method of claim 1 , wherein the host cell is in a mixed population of bacteria, the mixed population 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 the host cell, wherein the host cell is of a second bacterial species, wherein the second bacterial species is a different species than the first bacterial species.