A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells.
Chinese hamster ovary (CHO) cells and dihydrofolate reductase (dhfr)/methotrexate (MTX) gene amplification system are routinely used to generate stable producer CHO cell clones in biopharmaceutical industries. The present study proposes a novel method by the co-amplification of the silencing vector targeted to dhfr gene for improvements of selecting high-producing clones in dhfr-deficient and wild-type CHO cells. Using the silencing vector also resulted in improving the stability of the recombinant protein expression in the absence of MTX in the CHO/dhFr(-) and wild-type CHO cells. This new method is proposed to generate highly expressed stable cell clones of both dhfr-deficient and wild-type CHO cells for recombinant antigen production. Utilization of the silencing vector designed in this study can improve antigen expression through dhfr-directed gene amplification in other dhfr-competent cell lines for vaccine development.
Hong
Willy W L
WW
Institute of Biotechnology, Department of Life Science, National Tsing-Hua University, Hsinchu 30013, Taiwan.
Wu
Suh-Chin
SC
D000818:Animals; D000941:Antigens / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D016466:CHO Cells; D002999:Clone Cells; D006224:Cricetinae; D003412:Cricetulus; D005784:Gene Amplification; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins; D008727:Methotrexate / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D034622:RNA Interference; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics*; D014162:Transfection
D000941:Antigens; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D013762:Tetrahydrofolate Dehydrogenase; D008727:Methotrexate