| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | Enhancer sequences were synthesized and cloned into SpeI and NotI restriction sites between the loxP and the CMV core promoter (GenScript) to generate functional promoters. Features of both landing pads and mCherry RMCE donor plasmids are illustrated in Fig. 1a and detailed in Supplementary Tables S2 and S4. Fragments bearing msAb LC and HC genes or bsAb LC1, HC1, LC2 and HC2 were synthesized and cloned into a vector containing loxP and lox2272 sequences (GenScript). Various enhancer-CMV core promoters or full CMV promoter sequences were synthesized (GenScript) and cloned upstream of the LC or the HC genes, respectively. All constructs were validated by Sanger sequencing (GenScript) and were purified using NucleoBond Xtra Midi EF (Macherey-Nagel). | Get A Quote |
Constructing expression cell lines using site-specific integration (SSI) technology has become a prominent focus in cell line development (CLD) because of its potential to minimize clonal variation. However, this technology has not yet been widely adopted in commercial biopharmaceutical manufacturing. A major reason is the low yield of SSI-derived cell lines. To improve yield, we designed landing pad locus-specific synthetic promoters by screening for potent transcription factor regulatory elements (TFREs) within an SSI platform. We then combined these TFREs to create novel, strong promoters. All synthetic promoters exhibited reporter expression comparable to that of the CMV promoter, with some up to 20% strong... More
