Stable Cell Line Development Service –
Inducible expression is a valuable tool for functional analysis of assay systems with toxic targets. Also, adaptation of the signaling pathway is prevented when using inducible expression systems. Our ExoINDUCE technology combines the benefits of our ExoIN technology with the proven Tet/Dox system. This combination raises the Tet/Dox induction system to a new quality level, allowing tight regulation and high level expression upon induction.
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).
HEK293 ExoINDUCE-GFP cell pools
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.
HEK293 ExoINDUCE-GFP clones
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.
Main Advantages of trenzyme’s ExoINDUCE Technology
- Save time and money with our “all-in-one system” – only one round of cell line generation and clone screening is required to obtain the final inducible cell line
- Continuous selection of regulator expression on protein level (no silencing / no loss of regulator possible)
- Beneficial when selecting sensitive cell lines as only one antibiotic is needed for selection
- No licence fee and no royalties are due for generation and further use of your stable inducible cell line*
- ExoINDUCE technology is applicable to any cell line and all target classes
- Generation of ready-to-use stable assay cell lines within weeks – depending on target and parental cell line no single cell cloning is needed
- Clones with different target expression levels can be selected upon limiting dilution of the parental ExoINDUCE cell pool
“We were astonished by Trenzyme’s recently delivered custom ExoIN cell lines. Even “difficult-to-express” constructs show stable and robust expression. I highly recommend Trenzyme’s cell line development service.”
Process of Stable ExoINDUCE Cell Line Development with Inducible Target Expression
The generation process of an inducible cell line is divided into generation of a stable inducible cell pool (upper process line) and the generation of inducible clonal cell lines (lower process line). Directly after verification of the target expression and selection of a stable cell pool, single cell cloning can be started, if desired.
* if not otherwise agreed, for research and development purpose only
Inducible Expression FAQs
What is the benefit of the ExoINDUCE system in comparison to other inducible systems?
The ExoINDUCE system works as an “one plasmid system” and all elements for the generation of an inducible cell line are located on one plasmid. Due to the elaborated design of the system and the proven selection strategy, there is no need to generate a regulator cell line first – the final inducible cell line can be generated in only one step saving a lot of time and money.
Which target classes have you already worked with?
We already worked with all different target classes: transmembrane receptors, channels & transporters, intracelllular proteins, secreted proteins and many other target classes.
Does trenzyme only offer the generation of stable, inducible cell lines using the ExoINDUCE system?
No, if you provide your preferred parental cell line and your inducible expression system of choice, we will generate the recombinant cell line according to your needs.
Antje Fuhrmann, PhD
Application & Sales Manager
We would be happy to provide you with support on your cell line research project. Let us know your questions and requests, our scientific experts will reply shortly.
Get In Contact With Our Scientific Experts