Tet-On Advanced IRES Fluorescent Vector Set

Description

The Tet-On Advanced IRES Fluorescent Vector Set allows you to establish a tightly regulated and highly responsive inducible system for simultaneous coexpression of your gene of interest and a fluorescent protein marker. The system is turned on by adding doxycycline to the culture medium. The pTet-DualON Vector expresses the Tet-ON Advanced transactivator along with the green fluorescent protein, ZsGreen1, to allow you to sort for high-expression of the transactivator in target cells. With pTRE-Dual2, Tet-inducible coexpression of your gene of interest and the red fluorescent protein, mCherry, is controlled by the TRE-based promoter, PTight. Coexpression of two genes from a single transcription unit in each of these vectors is made possible by an internal ribosome entry site IRES2 that separates the two coding sequences. pTRE-Dual2-Luc is an inducible control vector that coexpresses mCherry and luciferase. To select stable cell lines, the vectors must be cotransfected with one of the provided linear selection markers. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Tetracycline-inducible expression of cytotoxic proteins.The Tet-On Advanced system was used to generate a double-stable HEK 293 cell line capable of induced expression of procaspase-8, a protein that plays a central role in apoptosis. Cells were treated for 8 hr with or without Dox and visualized by phase contrast microscopy. Due to the tight control of the caspase-8 transgene in the absence of Dox, the cells in Panel B appear very similar to the parental line Panel A . Apoptosis was only apparent after treatment of the cell line with Dox to induce expression of procaspase 8 Panel C . Back Monitor inducible expression using mCherry.pTRE-Dual vectors are supplied with the Tet-On Advanced IRES Fluorescent Vector Set Cat. # 631112 and Tet-Off Advanced IRES Fluorescent Vector Set Cat. # 631113 and provide simultaneous inducible expression of your transgene and our mCherry fluorescent protein from an IRES bicistronic mRNA. When your gene is turned on, so is mCherry, and your cells fluoresce red. Back IRES fluorescent vector sets Cat. # 631112, 631113 .The Tet-Dual transactivator expression vectors constitutively express either the Tet-On Advanced or Tet-Off Advanced protein, and the fluorescent protein, ZsGreen1 Panel A . pTRE-Dual2 is a Tet-regulated inducible expression vector that coexpresses mCherry and a second gene that is cloned in the multiple cloning site MCS Panel B . pTRE-Dual1 Cat. # 631114 coexpresses two user-defined genes. Back IRES fluorescent vector sets Cat. # 631112, 631113 .A 14-day old, puromycin-resistant colony of HEK 293 cells cotransfected with pTET-DualOFF, pTRE-Dual2-luciferase, and a linear puromycin selection marker exhibits dual fluorescent colors Panel A . The clone expresses Tet-Off Advanced protein and is green due to coexpressed ZsGreen1. In the absence of Dox, the cells are also red because Tet-Off Advanced protein activates the coexpression of luciferase and mCherry. The inducible expression of luciferase activity in this clone was measured and is shown in Panel B..