A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells

Abstract 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.