GAS Luciferase Reporter Lentivirus (IFN-γ/JAK/STAT1 Pathway)
Catalog #
78653
$835*
●
Purchase
●
Description
The GAS Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase gene driven by three copies of the interferon gamma (IFN-γ) activated sites (GAS) located upstream of the minimal TATA promoter (Figure 1) and a puromycin selection gene for the selection of stable clones. After transduction, the GAS-regulated gene expression in the target cells can be monitored by measuring the luciferase activity.
Figure 1. Schematic of the lenti-vector used to generate the GAS luciferase reporter lentivirus.
The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
Background
Interferon gamma (IFN-γ) is a pleiotropic cytokine with anti-viral, anti-tumor, and immune-modulatory functions. Cellular responses to IFN-γ are activated through its interaction with a heterodimeric receptor consisting of two subunits, Interferon gamma receptor 1 (IFNGR1) and Interferon gamma receptor 2 (IFNGR2), associated with kinases JAK1 and JAK2, respectively. Upon binding to this receptor, IFN-γ triggers JAK/STAT signaling. The activated STAT1 homodimers translocate to the nucleus where they bind interferon-gamma-activated sites (GAS) in the promoter of IFN-γ inducible genes, which stimulates IFN-γ-specific activation of gene transcription.
Storage/Stability
Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.
Applications
Screen for activators or inhibitors of IFN-γ-induced signal transduction pathways.
Generate GAS Luciferase Reporter stable cell line (puromycin resistant).
Shipping Temperature
-80°C
Notes
To generate a GAS luciferase reporter stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin for antibiotic selection of transduced cells.
The following Lentivirus Reporter Controls are available from BPS Bioscience to meet your experimental needs:
Negative Control Luciferase Lentivirus (BPS Bioscience #79578): Ready-to-transduce lentiviral particles expressing firefly luciferase under the control of a minimal promoter. The negative control is important to establish the specificity of any treatments and to determine the background reporter activity.
Renilla Luciferase Lentivirus (BPS Bioscience #79565): Ready-to-transduce lentiviral particles expressing Renilla luciferase under the CMV promoter. The Renilla Luciferase lentivirus can serve as an internal control to overcome sample-to-sample variability when performing dual-luciferase reporter assays.
Firefly Luciferase Lentivirus (BPS Bioscience #79692-G, #79692-H, #79692-P): Ready-to-transduce lentiviral particles expressing firefly luciferase under the CMV promoter. It serves as a positive control for transduction optimization studies.
Biosafety The lentiviruses are produced with SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
Powered by Bioz
