Engineering Chinese Hamster Ovary (CHO) cells for producing recombinant proteins with simple glycoforms by zinc-finger nuclease (ZFN)—mediated gene knockout of mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (Mgat1)

Mammalian cell-produced therapeutic proteins: heterogeneity derived from protein degradation

“…Additionally, with the introduction of new tools like zinc finger nucleases that allow easy and rapid manipulation of the host cell genome [for an example see Ref. [64 ]] and the recent public access to the CHO genome database [65 ], it is possible to tailor the host cell to meet product-specific quality requirements.…”

The GalNAc-type O-Glycoproteome of CHO Cells Characterized by the SimpleCell Strategy

“…The first successful design engineering of glycosylation in CHO was performed by two rounds of highly laborious and time consuming homologous recombinations to knockout FUT8 (37), but with the emerging precise gene editing technologies the efforts required have been drastically reduced as demonstrated by fast knock out of FUT8 with ZFN-mediated gene targeting (38). More recently, MGAT1 was knocked out in CHO GS cells to design a CHO platform for production of N-glycoproteins with homogeneous high mannose N-glycosylation, which is desirable for crystallization studies (39). The emergence of the precise gene editing technologies is expected to have major impact on custom design of glycosylation capacities of CHO and other cells (40).…”

Overexpression of Serpinb1 in Chinese hamster ovary cells increases recombinant IgG productivity

“…From the other half cell lysates were prepared for Western Blot analysis. The relative expression of Serpinb1 was then determined by SyBr Green using the primers listed in Supplementary For the plasmid stable pools and clones derived from transfectants of 666, total RNA isolation and Taqman ® qRT-PCR procedures were described in detail elsewhere (Sealover et al, 2013). The cycling conditions were the standard two-step Taqman ® conditions.…”