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Use of Flp-Mediated Cassette Exchange in the Development of a CHO Cell Line Stably Producing Erythropoietin  

Kim, Min-Soo (Department of Biological Sciences, Korea Advanced Institute of Science and Technology)
Lee, Gyun-Min (Department of Biological Sciences, Korea Advanced Institute of Science and Technology)
Publication Information
Journal of Microbiology and Biotechnology / v.18, no.7, 2008 , pp. 1342-1351 More about this Journal
Abstract
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 exchange.
Keywords
Flp-mediated cassette exchange; stable parental clone; long-term culture; EPO-producing CHO cell line;
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