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Use of Flp-mediated cassette exchange in the development of a CHO cell line stably producing erythropoietin
- Use of Flp-mediated cassette exchange in the development of a CHO cell line stably producing erythropoietin
- Kim, Min Soo; Lee, Gyun Min
- DGIST Authors
- Kim, Min Soo
- Issue Date
- Journal of Microbiology and Biotechnology, 18(7), 1342-1351
- Article Type
- Flp-mediated cassette exchange; stable parentalclone; long-term culture; EPO-producing CHO cell line
- The feasibility of the use of Flp-mediated cassette exchange in the development of a CHO cell line, which produces erythropoietin (EPO) stably and largely, was investigated. A stable, high enhanced green fluorescence protein (EGFP)-producing clone was screened by extensive flow cytometric analysis. An EPO expression unit was targeted into the premarked locus of the stable parental clone by Flp-mediated cassette exchange and a correctly targeted clone (FC28T7) was obtained. The EPO production of FC28T7 was proven to be stable in long-term culture. Furthermore, the Flp-mediated cassette exchange did not alter the stable parental clone's characteristics concerning transgene expression level and stability. Taken together, the data obtained here indicated that the establishment of CHO cell lines stably producing a desired protein is achievable using Flp-mediated cassette exchang. © The Korean Society for Microbiology and Biotechnology.
- Korean Society for Microbiology and Biotechnology
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