What is the Tet-Off and Tet-on systems used for?
The Tet-Off system allows silencing of gene expression by administration of tetracycline (Tc) or tetracycline-derivatives like doxycycline (dox), whereas the Tet-On system allows activation of gene expression by dox.
How does Tet-On system work?
Tet-On system: tetracycline binds the rtTA transcription factor and allows it to bind DNA at the promoter. Gene expression is induced in the presence of tetracycline.
How does DOX inducible system work?
We here review how a dox-controlled HIV-1 variant was designed and used to greatly improve the activity and dox-sensitivity of the rtTA transcriptional activator component of the Tet-On system.
How does tetracycline inducible system work?
A TRE is 7 repeats of a 19 nucleotide tetracycline operator (tetO) sequence, and is recognized by the tetracycline repressor (tetR). In the endogenous bacterial system, if tetracycline, or one of its analogs like doxycycline, are present, tetR will bind to tetracycline and not to the TRE, permitting transcription.
What is tetO promoter?
The TRE is made up of Tet operator (tetO) sequence concatemers fused to a minimal promoter, (commonly the minimal promoter sequence derived from the human cytomegalovirus (hCMV) immediate-early promoter). In the absence of Tc or Dox, tTA binds to the TRE and activates transcription of the target gene.
What is doxycycline inducible?
The doxycycline (dox)-inducible Tet-On system is widely used to control gene expression in mammalian cells. This system is based on the bacterial Tet operon, which has been modified and improved for its function in eukaryotic cells.
How is doxycycline different from tetracycline?
Doxycycline is better absorbed and reaches peak serum concentrations more quickly than tetracycline, but the difference isn’t large. Both drugs can be taken on an empty stomach but should be taken with food if there is gastrointestinal irritation as a result.
Is doxycycline in the same family as tetracycline?
The tetracycline family includes tetracycline, doxycycline and minocycline, all of which have been used as antibiotics effectively for decades.
What concentration of doxycycline induces induction?
50ng/ml doxycycline respectively (10ml final volume). Add 20μl OptiMEM to the 0ng/ml doxycycline wells. ✓ Incubate for 48-72 hrs to induce the expression of the gene of interest.