Applied Microbiology and Biotechnology, Vol.88, No.2, 425-436, 2010
Methods in mammalian cell line engineering: from random mutagenesis to sequence-specific approaches
Due to the increasing demand for recombinant proteins, the interest in mammalian cell culture, especially of Chinese hamster ovary cells, grows rapidly. This is accompanied by the desire to improve cell lines in order to achieve higher titers and a better product quality. Until recently, most cell line development procedures were based on random integration and gene amplification, but several methods for targeted genetic modification of cells have been developed. Some of those are homologous recombination, RNA interference and zinc-finger nucleases. Especially the latter two have evolved considerably and will soon become a standard for cell line engineering in research and industrial application. This review presents an overview of established as well as new and promising techniques for targeted genetic modification of mammalian cells.
Keywords:Cell line engineering;siRNA;miRNA;Zinc-finger nuclease (ZFN);Chinese hamster ovary (CHO) cells