DESCRIPTION

• Notes: To generate a Firefly Luciferase-mCherry stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of hygromycin (as pre-determined from a killing curve, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, following by clonal selection. BiosafetyThe lentiviruses are produced with a 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 further questions, please email support@bpsbioscience.com.
• Shiptemp: -80°C
• Warnings: Avoid freeze/thaw cycles
• Background: Firefly luciferase has been used as a sensitive reporter to study a wide range of biological responses. mCherry is a monomeric red fluorescent protein derived from DsRed found in the sea anemones Discosoma. It belongs to the mFruit family of monomeric red fluorescent proteins, which are improved versions of mRFP1 (monomeric red fluorescent protein 1) in terms of brightness and photostability. The use of fluorescent proteins allows for direct visualization of transfected or transduced cells under a fluorescent microscope or analysis by flow cytometry. The use of lentiviruses to introduce both luciferase and mCherry is a convenient strategy that allows expression of the markers in almost all mammalian cells and to easily determine transduction efficiency and access cellular responses.
• Description: Firefly Luciferase-mCherry Lentivirus (Hygromycin or Puromycin) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These viruses contain firefly luciferase and mCherry (Luc2-P2A-mCherry) driven by an EF1a promoter, and a hygromycin or puromycin selection marker (Figure 1). Luciferase and mCherry are co-expressed under the EF1A promoter in cells, allowing for flexibility in the detection of transduced cells. Figure 1. Schematic of the lenti-vector used to generate the Firefly Luciferase-mCherry Lentivirus (Puromycin). This is a SIN vector. Figure 1. Schematic of the lenti-vector used to generate the Firefly Luciferase-mCherry Lentivirus (Hygromycin). This is a SIN vector.
• Formulation: The lentivirus particles were produced in HEK293T cells in 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: Positive control in experiments involving transduction. Optimize transduction assays and track reporter protein expression over time. Generate cell pools or stable cell lines expressing firefly luciferase and mCherry following hygromycin or puromycin selection
• Product Type: Lentivirus
• Biosafety Level: BSL-2
• Related Products: 79565, 79692, 79980
• 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.
• Scientific Category: Cell Signaling Pathway