The Non-secreted Gaussia Luciferase Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles constitutively express Gaussia luciferase lacking the N-terminal signal peptide under a CMV promoter (Figure 1).
Figure 1. Schematic of the lenti-vector used to generate the Non-secreted Gaussia Luciferase Lentivirus
Two vials (500 μl x 2) of Non-secreted Gaussia Luciferase Lentivirus at a titer ≥5 x 106 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Materials Required But Not Supplied
• CHO-K1 cells (ATCC #CCL-61) • CHO growth medium or use BPS Thaw Medium 3 (BPS Bioscience #60186) • Polybrene (Millipore, #TR-1003-G) • 96-well tissue culture treated white clear-bottom assay plate (Corning, #3610) • Coelenterazine native (NanoLight Technology, #303-500): Make a 2 mM stock in Ethanol • CHO lysis buffer: PBS+1% TritonX-100 • Luminometer
Formulation
The lentiviruses were produced from HEK293T cells in medium containing 90% DMEM + 10% FBS.
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.
Instructions for Use
See assay protocol for detailed instructions.
Shipping Temperature
-80°C
Warnings
Biosafety
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. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.