MRE Luciferase Reporter Lentivirus (Heavy Metal Stress)

Catalog #

78910

$875 *

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

MRE Luciferase Reporter Lentiviruses (Heavy Metal Stress) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These viruses contain a firefly luciferase reporter driven by multiple copies of MRE (metal response elements) located upstream of the minimal TATA promoter, and a puromycin selection marker (Figure 1).

After transduction, transcription mediated by MRE can be monitored by measuring luciferase activity.

Figure 1. Schematic of the lenti-vector used to generate the MRE Luciferase Reporter Lentivirus (Heavy Metal Stress).

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Product Data Gallery

Activation of MRE luciferase reporter activity in HepG2 cells transduced with MRE Luciferase Reporter Lentivirus (Heavy Metal Stress).

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
HepG2 Cells	ATCC #HB-8065
Thaw Medium 1	BPS Bioscience #60187
Assay Medium 1	BPS Bioscience #78674
ZnCl2	Sigma #Z0152
96-well tissue culture, clear-bottom, white plate	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. Virus particles can be packaged in custom formulations by special request, for an additional fee.

Background

Metal ions are important structural components of many enzymes and cofactors of enzymatic reactions. Essential metals include zinc, copper and iron. Zinc is a common co-factor of hydrolytic enzymes and a structural factor in proteins like zinc finger proteins, known to bind to more than 300 enzymes and 2000 transcription factors. It participates in responses to oxidative stress, DNA replication and repair, cell cycle, apoptosis and aging. It is crucial for the proper development of the brain, and tight control of its levels in the central neural system is essential. In vertebrates, zinc homeostasis is kept via the zinc-dependent regulation of the transcription of metallothioneins (MTs), scavengers of heavy metals that protect against oxidative stress, by the interaction of MTF-1 (MRE-binding transcription factor 1) with MREs (metal response element). MRE is the cis-acting DNA element that can be found in promoters of zing-regulated genes, and that binds MTF-1 and other zinc-related transcription factors. Dysregulation of metal levels in the organism can lead to pathologies such as Alzheimer’s disease, with excess zinc linking to hyperphosphorylated tau aggregation. An understanding of the pathways and partners involved in metal stress regulation will bring benefits to patients suffering from zinc-related diseases.

References

Giedroc D., et al., 2001 Antioxidants &Redox Signaling 3(4): 577-596.
Li Z., et al., 2023 Biomolecules 13(1):28.

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.

Growth Media

For best results, the use of validated and optimized media from BPS Bioscience is highly recommended. Other preparations or formulations of media may result in suboptimal performance.

Media Required for the Proposed Assay
Thaw Medium 1 (BPS Bioscience #60187): MEM medium supplemented with 10% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin.

Assay Medium 1C (BPS Bioscience #78674): MEM medium supplemented with 0.5% charcoal stripped FBS, 1% non-essential amino acids, 1 mM Na pyruvate.

Applications

Generate cell pools or stable cell lines expressing the MRE luciferase reporter following puromycin selection.

Shipping Temperature

-80°C

Notes

To generate a MRE 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 (as pre-determined from a killing curve, FAQs (bpsbioscience.com) for antibiotic selection of transduced cells, followed by clonal selection.
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. This 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 control of a CMV promoter. The Renilla luciferase lentiviruses 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 control of a CMV promoter. It serves as a positive control for transduction optimization studies.

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

Biosafety
The lentiviruses are produced with SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and 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.

Warnings

Avoid freeze/thaw cycles

Scientific Category

Metabolic Enzymes

Regulatory Status

Research Use Only

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MRE Luciferase Reporter Lentivirus (Heavy Metal Stress)

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