Bacterial artificial chromosome library for genome-wide analysis of Chinese hamster ovary cells

Generation of monoclonal antibody-producing mammalian cell lines

“…With the approval of a glycomodified mAb from an engineered CHO cell line, and with over ten others under testing, we will likely see an increase in such products in the future [66]. Increased knowledge of the CHO genome [28,74,75], transcriptome [69,76] and proteome [77] would allow greater understanding of the complex interactions taking place for improved cell engineering approaches. Construction of dynamic computational models has worked well in the production of microbial cells and this extra information will eventually allow similar implementation in mammalian cells [39,78,79].…”

3.2 Chromosome Rearrangements and Gene Amplification

“…The head-to-head DHFR amplicon in CHOC 400 has been reported, and its structure generally supports the idea of sister chromatid fusion [39,40]. The authors also investigated the genomic structure of the exogenous DHFR amplicon in the CHO DR1000L-4N cell line from the construction of the CHO BAC library [19,41] (Figure 3.2.10). They determined the complete nucleotide sequence of the insert in the Cg0031N14 BAC clone, which contains the amplicon, by shotgun sequencing.…”

“…These approaches could greatly contribute to the genome-scale reconstruction of industrial mammalian cells. Previously, the authors constructed the first genomic BAC library from the geneamplified CHO cell (CHO DR1000L-4N) genome for genome-wide analysis [19]. The BAC clones of this library could be landmarks for a physical map of the CHO cell genome, which would be essential for the basic research and industrial applications of the CHO cell genome.…”

Ubiquitous Chromatin Opening Elements (UCOEs) effect on transgene position and expression stability in CHO cells following methotrexate (MTX) amplification

“…Various studies have suggested that the location of integrated plasmid DNA in the CHO cell chromosome plays a role in productivity and stability . Recently, BAC libraries were used to investigate the chromosomal region adjacent to the transgene vector in amplified CHO cell lines . It has been suggested that endogenous repetitive structures of the amplified region on a specific CHO chromosome may promote gene amplification and increase the stability of the inserted gene .…”

“…Recently, BAC libraries were used to investigate the chromosomal region adjacent to the transgene vector in amplified CHO cell lines . It has been suggested that endogenous repetitive structures of the amplified region on a specific CHO chromosome may promote gene amplification and increase the stability of the inserted gene . It is of particular interest in mammalian cells to examine the role of the integration site in amplification and genomic stability.…”