Pathway integration and expression in host cells
Abstract
Provided herein are methods for integrating a gene of interest into a chromosome of a host cell. In some embodiments, the methods include introducing into a host cell a first plasmid comprising a transposase coding sequence and a donor sequence, which includes a selectable marker coding sequence flanked by a first and a second lox site and is itself flanked by inverted repeats recognized by the transposase. Following transposase-mediated chromosomal integration of the donor sequence into the host cell, a second plasmid is introduced, which comprises the gene of interest and a second selectable marker coding sequence, both flanked by a first and a second lox site. The gene of interest is chromosomally integrated into the host cell by recombinase-mediated cassette exchange (RMCE) between the donor sequence and the second plasmid via Cre-/cuc recombination. Further provided herein are host cells, vectors, and methods of producing a product related thereto.
- Inventors:
- Issue Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1998473
- Patent Number(s):
- 11674145
- Application Number:
- 16/072,174
- Assignee:
- The Regents of the University of California (Oakland, CA)
- DOE Contract Number:
- AC02-05CH11231
- Resource Type:
- Patent
- Resource Relation:
- Patent File Date: 01/24/2017
- Country of Publication:
- United States
- Language:
- English
Citation Formats
Yoshikuni, Yasuo, Wang, Gaoyan, Zhao, Zhiying, Coates, Robert Cameron, Cheng, Jan-Fang, and Robinson, David. Pathway integration and expression in host cells. United States: N. p., 2023.
Web.
Yoshikuni, Yasuo, Wang, Gaoyan, Zhao, Zhiying, Coates, Robert Cameron, Cheng, Jan-Fang, & Robinson, David. Pathway integration and expression in host cells. United States.
Yoshikuni, Yasuo, Wang, Gaoyan, Zhao, Zhiying, Coates, Robert Cameron, Cheng, Jan-Fang, and Robinson, David. Tue .
"Pathway integration and expression in host cells". United States. https://www.osti.gov/servlets/purl/1998473.
@article{osti_1998473,
title = {Pathway integration and expression in host cells},
author = {Yoshikuni, Yasuo and Wang, Gaoyan and Zhao, Zhiying and Coates, Robert Cameron and Cheng, Jan-Fang and Robinson, David},
abstractNote = {Provided herein are methods for integrating a gene of interest into a chromosome of a host cell. In some embodiments, the methods include introducing into a host cell a first plasmid comprising a transposase coding sequence and a donor sequence, which includes a selectable marker coding sequence flanked by a first and a second lox site and is itself flanked by inverted repeats recognized by the transposase. Following transposase-mediated chromosomal integration of the donor sequence into the host cell, a second plasmid is introduced, which comprises the gene of interest and a second selectable marker coding sequence, both flanked by a first and a second lox site. The gene of interest is chromosomally integrated into the host cell by recombinase-mediated cassette exchange (RMCE) between the donor sequence and the second plasmid via Cre-/cuc recombination. Further provided herein are host cells, vectors, and methods of producing a product related thereto.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2023},
month = {6}
}
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