The Myc 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 the Myc response element located upstream of the minimal TATA promoter (Figure 1) and an antibiotic selection gene (puromycin) or a G418 selection marker for the selection of stable clones. After transduction, the Myc signaling pathway in the target cells can be monitored by measuring the luciferase activity.
Figure 1. Schematic of the lenti-vector used to generate the Myc luciferase reporter lentivirus. (Puromycin)
Figure 1: Schematic of the lenti-vector used to generate Myc Luciferase Reporter Lentivirus. (G418)
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Inhibition of Myc luciferase reporter activity by ICG-001 in HCT116 cells transduced with Myc Luciferase Reporter Lentivirus. (Puromycin)
Inhibition of Myc luciferase reporter activity by ICG-001 in HCT116 cells transduced with Myc Luciferase Reporter Lentivirus. (G418)
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 and produced at higher titers by special request, for an additional fee.
Background
The Myc signaling pathway plays an important role in cell proliferation, differentiation, transformation and apoptosis. c-Myc is a transcription factor that heterodimerizes with MAX to regulate Myc signaling pathway-responsive genes. Myc can be activated by the Wnt/β-catenin pathway. Genetic alterations in MYC have been linked to a number of human cancers, including Burkitt’s lymphoma, cervical, ovarian, breast, lung, and pancreatic carcinomas. Thus, Myc is a promising therapeutic target for cancer treatment.
References
Pelengaris S., et al., 2002 Nat. Rev. Cancer. 2(10): 764-76.
Storage/Stability
Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the virus at -80°C for up to 12 months from date of receipt. Avoid repeated freeze-thaw cycles. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.
Growth Media
For best results, it is highly recommended to use these validated and optimized media from BPS Bioscience. Other preparations or formulations of media may result in suboptimal performance.
Thaw Medium 7 (BPS Bioscience, #60185): McCoy’s 5A medium with 10% FBS, 1% Penicillin/Streptomycin.
Assay Medium 7B (BPS Bioscience, #79718): Opti-MEM I + 0.5% FBS + 1% non-essential amino acids + 1 mM sodium pyruvate + 1% penicillin/ streptomycin.
Applications
Screen for activators or inhibitors of the Myc signaling pathway in transduced target cells.
Generate cell pools or stable cell line expressing the Myc luciferase reporter following G418 or puromycin selection
Shipping Temperature
-80°C
Notes
To generate a Myc Luciferase reporter-expressing 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, https://bpsbioscience.com/cell-line-faq), 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 (#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 (#79565): Ready-to-transduce lentiviral particles expressing Renilla luciferase under the control of a CMV promoter. The RLuc lentivirus can serve as an internal control to overcome sample-to-sample variability when performing dual-luciferase reporter assays.
Firefly Luciferase Lentivirus (#79692-G, #79692-H, #79692-P): Ready-to-transduce lentiviral particles expressing firefly luciferase under the control of a CMV promoter. It can serve as a positive control for transduction optimization studies.
Biosafety The lentiviruses are produced with the third generation 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.
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