Tet Inducible Gene Expression Stable Cell Line

VectorBuilder can custom build Tet inducible gene expression stable cell lines with minimal background expression and high induction of your gene of interest (GOI). Additionally, we can engineer stable cell lines expressing Tet regulatory protein (e.g. rtTA, tTS/rtTA, tet3G, etc.) which can then be transduced/transfected with viruses/plasmids carrying your GOI driven by TRE for flexible experimental design and reliable Tet inducible gene expression. For Tet-On stable cell lines, induction of the GOI is validated by RT-qPCR. Additionally, a series of standard QC assays such as sterility tests and mycoplasma detection are performed before releasing the final cell line products.

Highlights

  • Switch-like gene activation: our vector system allows for tetracycline-regulated on-and-off switch for controlling gene expression, minimizing background expression and exhibiting high sensitivity. 
  • High-level expression: the TRE promoter can drive very high levels of gene expression in its induced state.
  • Rapid turnaround: engineered cells can be delivered in as fast as 9 weeks starting from vector design.

Service Details

Workflow Tet inducible gene expression stable cell line engineering

Figure 1. Typical workflow of Tet inducible gene expression stable cell line production.

Price and turnaround
Service TypeDeliverablePrice (USD)*Turnaround
Tet-inducible gene expressionMixed pool (>106 cells/vial, 2 vials)From $3,9999-15 weeks
Two single clones (>106 cells/vial, 2 vials per clone)From $4,99914-20 weeks

* Additional charge will apply for extra single clones or vials.

QC assays
AssaysMethods
Tet-induced overexpression validation (default)RT-qPCR
Expression test (add-on)WB, IF, FACS
Sterility (default)PCR for mycoplasma detection, bioburden test for sterility detection 

Downstream services

We can perform various phenotypic assessments and functional validation of your engineered cell lines, including assays for proliferation, apoptosis, migration, viability, cytotoxicity, etc. 

Case Studies

Tet inducible EGFP expression stable cell line validation

Figure 2. Tet-inducible EGFP expression in 293T cells. The tTS/rtTA stable expression 293T cells were transduced with lentivirus carrying TRE-driven EGFP at an MOI of 1. EGFP expression was measured in the absence (Dox-) or in the presence (Dox+) of doxycycline (tetracycline derivative, 10 ng/ml) using (A) microscopy, (B) flow cytometry, and (C) western blot at 24 h and 48 h post-transduction. MFI, median fluorescence intensity.

FAQ

How is gene induction controlled in Tet-regulated systems?

The tetracycline inducible gene expression system is a powerful tool to control the timing of gene expression in mammalian cells. Our Tet-On vector systems are designed to achieve nearly complete silencing of a GOI in the absence of tetracycline, and strong, rapid expression in response to the addition of tetracycline. This is achieved through a multicomponent system which incorporates active silencing by the tTS protein in the absence of tetracycline and strong activation by the rtTA protein in the presence of tetracycline. In the absence of tetracycline, the tTS protein derived from the fusion of TetR (Tet repressor protein) and KRAB-AB (the transcriptional repressor domain of Kid-1 protein) binds to the TRE promoter, leading to the active suppression of gene transcription. The rtTA protein, on the other hand, derived from the fusion of a rTetR (a mutant Tet repressor) and VP16 (the transcription activator domain of virion protein 16 of herpes simplex virus), binds to the TRE promoter to activate gene transcription only in the presence of tetracycline. Our Tet-Off vector system can be used for inhibiting target gene expression in the presence of tetracycline regulated by the binding of the tTA protein to the TRE promoter.

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