US 11,896,626 B2
Multistrain population control systems and methods
Jeff Hasty, Encinitas, CA (US); Michael Julius Liao, San Diego, CA (US); and Muhammad Omar Din, San Diego, CA (US)
Assigned to The Regents of the University of California, Oakland, CA (US)
Appl. No. 16/972,405
Filed by The Regents of the University of California, Oakland, CA (US)
PCT Filed Jun. 7, 2019, PCT No. PCT/US2019/036179
§ 371(c)(1), (2) Date Dec. 4, 2020,
PCT Pub. No. WO2019/237083, PCT Pub. Date Dec. 12, 2019.
Claims priority of provisional application 62/682,755, filed on Jun. 8, 2018.
Prior Publication US 2021/0169941 A1, Jun. 10, 2021
Int. Cl. A61K 35/74 (2015.01); C07K 14/245 (2006.01); C12N 1/20 (2006.01); A61K 35/00 (2006.01)
CPC A61K 35/74 (2013.01) [C07K 14/245 (2013.01); C12N 1/20 (2013.01); A61K 2035/115 (2013.01)] 16 Claims
 
1. 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.