DESCRIPTION

• 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 support@bpsbioscience.com. BiosafetyThe 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.
• Shiptemp: -80°C (dry ice)
• Warnings: Avoid freeze/thaw cycles
• Category: Cell Signaling/Lentivirus
• 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.
• 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).
• 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.
• 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.
• Unspsc Code: 41106621
• Unspsc Name: Virus mediated expression vectors or kits
• Applications: Generate cell pools or stable cell lines expressing the MRE luciferase reporter following puromycin selection.
• Product Type: Lentivirus
• Biosafety Level: BSL-2
• Related Products: 79578, 79565, 79692
• Storage Stability: Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.
• Scientific Category: Metabolic Enzymes
• Signed License Required: No