	US 11,718,820 B2
	Genetically modified haploid Issatchenkia orientalis
Kenneth R. Finley, St. Bonifacius, MN (US); Holly Jessen, Chanhassen, MN (US); Erin Kathleen Marasco, Excelsior, MN (US); Thomas William McMullin, Minnetonka, MN (US); Ana Negrete-Raymond, Chanhassen, MN (US); and Amit Vas, Minneapolis, MN (US)
Assigned to CARGILL, INCORPORATED, Wayzata, MN (US)
Appl. No. 16/638,251
Filed by CARGILL, INCORPORATED, Wayzata, MN (US)
PCT Filed Aug. 2, 2018, PCT No. PCT/US2018/044998 § 371(c)(1), (2) Date Feb. 11, 2020, PCT Pub. No. WO2019/036202, PCT Pub. Date Feb. 21, 2019.
Claims priority of provisional application 62/546,662, filed on Aug. 17, 2017.
Prior Publication US 2020/0172855 A1, Jun. 4, 2020
Int. Cl. C12N 1/14 (2006.01); C12N 1/16 (2006.01); C07K 14/39 (2006.01); C12Q 1/02 (2006.01)

CPC C12N 1/16 (2013.01) [C07K 14/39 (2013.01); C12Q 1/025 (2013.01)]

15 Claims

1. A method of making a viable Issatchenkia orientalis organism that contains N to 2N−1 chromosomes, comprising the steps of:

a) growing parent diploid and/or tetraploid I. orientalis cells in the presence of an agent selected from the group consisting of nocodazole, benomyl, colchicine, para-fluoro-phenylalanine, and combinations thereof; and

b) isolating and identifying at least a portion of the viable daughter cells that contain N to 2N−1 chromosomes, wherein 2N represents the number of chromosomes in wild-type Issatchenkia orientalis.