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US 11,896,626 B2Granted
Multistrain population control systems and methods
AssigneeThe Regents of the University of California
InventorsJeff Hasty, Michael Julius Liao, Muhammad Omar Din
Life of this patent
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Abstract
Provided herein are multi-strain population control systems, methods, kits, and compositions. Also provided are methods, systems, kits, and compositions for culturing bacterial cells in multi-strain ecosystems, and temporally arranged multi-strain ecosystems or cultures using a synchronized lysis circuit in combination with multiple toxin/antitoxin systems to cycle continuously over a long period of time.
Claims16
A method of maintaining a bacterial co-culture, the method comprising:
- culturing a first bacterial strain for a first period of time in a growth environment;
- adding a second bacterial strain to the growth environment and culturing the second bacterial strain for a second period of time;
- adding a third bacterial strain to the growth environment and culturing the third bacterial strain for a third period of time,
- wherein each of the first, second, and third bacterial strains comprises a toxin system;
- wherein the toxin system of the first bacterial strain produces a first toxin/first antitoxin pair and a third antitoxin from a third toxin/third antitoxin pair wherein the first bacterial strain does not produce the third toxin;
- wherein the toxin system of the second bacterial strain produces a second toxin/second antitoxin pair and the first antitoxin from the first toxin/first antitoxin pair wherein the second bacterial strain does not produce the first toxin;
- and
- wherein the toxin system of the third bacterial strain produces a third toxin/third antitoxin pair and the second antitoxin from the second toxin/second antitoxin pair wherein the third bacterial strain does not produce the second toxin, and
- wherein each of the first, second, and third bacterial strains comprises
- a lysis plasmid having a lysis gene under the control of an activatable promoter;
- and
- an activator plasmid having an activator gene, the expression of which promotes the accumulation of a quorum-sensing molecule,
- wherein both the activatable promoter of the lysis gene and the expression of the activator gene is activated by the quorum-sensing molecule.
10 more elements
A method of maintaining a bacterial co-culture, the method comprising:
- providing n bacterial strains comprising at least a first bacterial strain, a second bacterial strain, and an nth bacterial strain that are distinct from each other;
- co-culturing each of the bacterial strains sequentially for one or more independent periods of time in a growth environment;
- wherein n is at least 3;
- wherein each of the n bacterial strains comprises a toxin system;
- wherein the toxin system of the first bacterial strain produces a first toxin/first antitoxin pair and an nth antitoxin from an nth toxin/nth antitoxin pair, wherein the first toxin is effective against the nth bacterial strain;
- wherein the toxin system of the second bacterial strain produces a second toxin/second antitoxin pair and the first antitoxin from the first toxin/first antitoxin pair, wherein the second toxin is effective against the first bacterial strain;
- and
- wherein the toxin system of the first bacterial strain produces an nth toxin/nth antitoxin pair and an n−1th antitoxin from an n−1th toxin/n−1th antitoxin pair, wherein the nth toxin is effective against the n−1th bacterial strain.
5 more elements
Dependencies are inferred from the claim text.