	US 12,134,764 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)
Filed by CARGILL, INCORPORATED, Wayzata, MN (US)
Filed on Jun. 15, 2023, as Appl. No. 18/335,717.
Application 18/335,717 is a division of application No. 16/638,251, granted, now 11,718,820, previously published as PCT/US2018/044998, filed on Aug. 2, 2018.
Claims priority of provisional application 62/546,662, filed on Aug. 17, 2017.
Prior Publication US 2024/0018464 A1, Jan. 18, 2024
Int. Cl. 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 viable Issatchenkia orientalis cell that contains N to 2N−1 chromosomes, wherein 2N represents the number of chromosomes in wild-type Issatchenkia orientalis, and comprises at least one exogenous gene that encodes for a gene product, the at least one exogenous gene integrated into at least one chromosome, wherein the cell is made by a method comprising the steps of:

a) growing a parent diploid and/or tetraploid I. orientalis cell 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.