US 12,077,786 B2
Methods of cell selection and modifying cell metabolism
Robert Young, Visp (CH); Christopher Mark Smales, Visp (CH); Colin Mark Jaques, Visp (CH); Andrew J. Racher, Visp (CH); Gurdeep Singh, Visp (CH); James Budge, Visp (CH); and Joanne Robool, Visp (CH)
Assigned to LONZA LTD, Visp (CH)
Appl. No. 16/966,768
Filed by LONZA LTD, Visp (CH)
PCT Filed Feb. 1, 2019, PCT No. PCT/US2019/016403
§ 371(c)(1), (2) Date Jul. 31, 2020,
PCT Pub. No. WO2019/152876, PCT Pub. Date Aug. 8, 2019.
Claims priority of provisional application 62/625,773, filed on Feb. 2, 2018.
Prior Publication US 2023/0174971 A1, Jun. 8, 2023
Int. Cl. C12N 9/06 (2006.01); C12N 9/02 (2006.01); C12N 15/10 (2006.01); C12N 15/63 (2006.01)
CPC C12N 9/0008 (2013.01) [C12N 15/1075 (2013.01); C12N 15/63 (2013.01)] 10 Claims
OG exemplary drawing
 
1. A method of identifying, selecting or culturing a mammalian cell comprising a subject nucleic acid sequence, the method comprising:
a) providing a cell comprising a heterologous nucleic acid comprising:
(i) the subject nucleic acid sequence; and
(ii) a nucleic acid sequence encoding a pyroline-5-carboxylate synthase (P5CS) molecule; and
b) culturing the cell comprising the heterologous nucleic acid sequence in the presence of media having an insufficient level of proline to support growth of a cell not having elevated P5CS enzymatic activity that is the same cell type as in step a), under conditions sufficient to allow for growth of a cell comprising the heterologous nucleic acid sequence, wherein the media further comprises an inhibitor of enzymatic activity of the P5CS molecule and wherein the inhibitor is a proline analog,
thereby identifying, selecting or culturing, a cell comprising the heterologous nucleic acid sequence.