How does rtTA work?

How does rtTA work?

Like tTA, rtTA is a fusion protein comprised of the TetR repressor and the VP16 transactivation domain; however, a four amino acid change in the tetR DNA binding moiety alters rtTA’s binding characteristics such that it can only recognize the tetO sequences in the TRE of the target transgene in the presence of the Dox …

What is tetracycline transactivator?

Tet-Off and Tet-On The Tet-Off system makes use of the tetracycline transactivator (tTA) protein, which is created by fusing one protein, TetR (tetracycline repressor), found in Escherichia coli bacteria, with the activation domain of another protein, VP16, found in the herpes simplex virus.

What is DOX in biology?

A derivative of tetracycline, doxycycline (dox), is a preferred effector for tetracycline transregulation. Dox binds with high affinity to tTA and rtTA; thus, dox can be used in both tetracycline on and off systems.

What is transactivation activity?

In the context of gene regulation: transactivation is the increased rate of gene expression triggered either by biological processes or by artificial means, through the expression of an intermediate transactivator protein.

What does rtTA stand for?

rtTA

Acronym Definition
rtTA Reverse Tetracycline Transactivator (immunology)
rtTA Race to the Altar (reality TV)
rtTA Read the Title Again

What is transactivator in biology?

The transactivator gene expresses a transcription factor that binds to specific promoter region of DNA. By binding to the promoter region of a gene, the transcription factor causes that gene to be expressed.

What is a transactivator of transcription?

The transactivator of transcription (TAT) protein transduction domain is an 11-amino acid positively charged peptide that has been shown to pull diverse molecules across cell membranes in vitro and in vivo.

Is CMV a strong promoter?

While there is cell type to cell type variability for all the promoters, CMV promoter is the most variable, being very strong in some cell types (e.g., 293T and CMMT) and rather weak in others (e.g., MRC5 and MSC).

Does Doxy have gram-negative coverage?

Doxycycline has a broad spectrum of activity and is effective against gram-positive and gram-negative, aerobic and anaerobic bacteria, spirochetes, and mycoplasma.

What is a transactivation assay?

The principle of in vitro androgen transactivation assay, based on stable transfection of a cell line with two plasmids; one encoding the androgen receptor and the other, the androgen response element (ARE) upstream of a reporter (REP) gene such as luciferase.

What is Transactivator in biology?

What is the CMV promoter used for?

The CMV promoter is a commonly used promoter for the production of high level recombinant protein in mammalian cells17. However, the expression level of the transgene driven by CMV promoter decreases with extended culture times because of transcriptional silencing, which is associated with DNA methylation18, 19.