US 12,168,769 B2
Methods for genomic integration in Pichia and other host cells
Hanxiao Jiang, Fremont, CA (US)
Assigned to Amyris, Inc., Emeryville, CA (US)
Filed by Amyris, Inc., Emeryville, CA (US)
Filed on Jun. 15, 2022, as Appl. No. 17/841,429.
Application 17/841,429 is a division of application No. 16/646,001, granted, now 11,390,874, previously published as PCT/US2018/050613, filed on Sep. 12, 2018.
Claims priority of provisional application 62/666,923, filed on May 4, 2018.
Claims priority of provisional application 62/560,026, filed on Sep. 18, 2017.
Prior Publication US 2022/0315937 A1, Oct. 6, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/81 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/815 (2013.01) [C12N 15/905 (2013.01)] 19 Claims
 
1. A method of producing an antibody, the method comprising,
culturing a yeast host cell comprising (i) a nucleic acid sequence encoding an antibody in the yeast host cell genome and (ii) a circular extrachromosomal nucleic acid comprising a coding sequence for a selectable marker, under conditions suitable for production of the antibody, wherein the host cell further comprises a nucleic acid that encodes an RNA-guided DNA endonuclease, wherein the nucleic acid is integrated in a YKU70 gene, thereby reducing non-homologous end joining (NHEJ) activity in the host cell as compared to a host cell in which YKU70 is intact; and
recovering the antibody produced by the yeast host cell.