Inducible Expression
based on unique ExoINDUCE technology
Our smart selection strategy enables highly effective inducible target protein expression in every mammalian cell line. trenzyme’s all-in-one ExoINDUCE system ensures development of stable inducible clonal cell lines with only one round of stable selection and clonal cell line generation. All features needed for cell line generation are encoded on one single plasmid: target, transcriptional regulator and antibiotic resistance marker.
The ExoIN system couples the expression of the transcriptional activator to the expression of the selection marker on protein level which results in a streamlined process based on only one antibiotic selection. Due to this linkage, the risk of silencing or loss of the regulator is marginal. The use of an activator as inducible system leads to low background expression levels and the opportunity of directly generating functional pools without clonal selection (see Figure 1). Additionally including clonal selection, a cell line with desired target expression level and even lower background expression can be generated (see Figure 2).
Figure 1: HEK293 parental and HEK293 ExoINDUCE-GFP pool.
HEK293 cells were transfected with ExoINDUCE all-in-one plasmid encoding GFP as target. A bulk pool was selected with hygromycin and analyzed 2 weeks after start of selection. For analysis, the bulk pool was induced by cultivation for 24h in medium containing 1 µg/mL doxycycline. 45.7 % GFP positive cells were detected by flow cytometry after induction.
Figure 2: HEK293 ExoINDUCE-GFP clones.
HEK293 ExoINDUCE-GFP clones were generated by limiting dilution of the bulk pool. Clones were analyzed 2 weeks after seeding and clones with different expression patterns were generated. For analysis, clones were induced by cultivation for 24h in medium containing 1 µg/mL doxycycline.
High, medium and low expressing clones are shown in histograms with and without doxycycline.