UAS Luciferase Reporter Lentivirus

Catalog #

78631

$835 *

Size: 500 µl x 2

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The UAS (Upstream Activation Sequence) Luciferase Reporter Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most mammalian cells, including primary and non-dividing cells. The particles contain a firefly luciferase reporter driven by a multimerized GAL4 upstream activation sequence (UAS) located upstream of the minimal TATA promoter. The lentiviruses also contain a puromycin selection marker (Figure 1). After transduction, the UAS-controlled signaling pathway in the target cells can be monitored by measuring luciferase activity.

Figure 1. Schematic of the lenti-vector used to generate the UAS luciferase reporter lentivirus.

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Product Data Gallery

Dexamethasone-induced activation of the UAS luciferase reporter with GAL4 DBD-GR in HEK293 cells transduced with UAS Luciferase Reporter Lentivirus and GAL4 DBR-GR Lentivirus

Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of lentivirus at a titer >10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
HEK293	ATCC #CRL-1573
Thaw Medium 1	BPS Bioscience #60187
GAL4 DBD-GR Lentivirus	BPS Bioscience #78632
Dexamethasone	Sigma #D4902
Polybrene Infection / Transfection Reagent	Millipore #TR-1003-G
96-well Clear Bottom White Polystyrene TC-treated	Corning #3610
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690
Luminometer

Formulation

The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.

Background

GAL4/UAS is a binary gene expression system used for modeling expression vectors or transgenic animals with GAL4 and UAS constructs. UAS is a regulatory sequence critical for the induction and the increased expression of a nearby gene of interest. GAL4 binds to UAS to activate transcription from a basal promoter located downstream of the activation sequence. Gal4/UAS is a highly advantageous system that can induce gene expression at a desired time and at higher levels than endogenous tissue-specific promoters. It can link a portion of a nuclear receptor to a reporter system to study ligand binding or to screen small molecules.

GAL4 DBD-GR lentivirus (BPS Bioscience #78632), expresses a fusion protein where the glucocorticoid receptor (GR) ligand binding domain is fused to the DNA binding domain of GAL4 (GAL4 DBD). After co-transduction of UAS luciferase reporter lentivirus and GAL4 DBD-GR lentivirus. The glucocorticoid-induced activation of the glucocorticoid receptor can be monitored by measuring the luciferase activity.

The glucocorticoid signaling pathway plays an important role in development, fluid homeostasis, cognition, immune response and metabolism. Glucocorticoids are a class of steroid hormones that bind to the glucocorticoid receptor, causing it to translocate to the nucleus. Upon translocation, the receptor can regulate the transcription of many genes, including those that regulate glucose metabolism and inflammatory responses.

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

In combination with GAL4 DBD-GR lentivirus (BPS Bioscience #78632) to monitor glucocorticoid signaling pathway activity.
Generate a UAS Luciferase Reporter stable cell line (puromycin resistant)

Shipping Temperature

-80°C

Notes

To generate the UAS 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 Luciferase Lentivirus Reporter Controls are available from BPS Bioscience to meet your experimental needs:
1. 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.
2. Renilla Luciferase Lentivirus (BPS Bioscience #79565-P, #79565-G): 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.
3. 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 the 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.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

Replication Incompetent

Regulatory Status

Research Use Only