University of California

Robert Cameron Coates

Yasuo Yoshikuni

Zhiying Zhao

Jan-Fang Cheng

David Robinson

Gaoyan Wang

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.

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Lafayette, CA, United States of America

Robert Cameron Coates