Our GFP Reporter Lentivirus contains replication-defective, 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 contain an EF1α promoter-driven GFP construct (Accession # NM001527), and confer both GFP expression and puromycin resistance to the target cells (Figure 1). GFP expression and transduction efficiency can easily be verified and optimized via fluorescence microscopy or flow cytometry.

Figure 1. Schematic of the lentiviral vector used to generate the GFP reporter

• BPS Medium 2 (BPS Bioscience #79704):

• RPMI 1640 medium (Thermo Fisher, #A1049101) supplemented with 10% FBS (Thermo Fisher, #26140079), 1mM Sodium pyruvate (Corning, #25-000-CL), 1% Non-essential amino acids (Corning, #25-025-CL), 1% Penicillin/streptomycin (Thermo Fisher, #15140122) T cell activation reagents (αCD3/αCD28 or Phytohaemagglutinin (PHA))

• Recombinant Human IL-2 (BPS Bioscience, #90184)
• Polybrene (Millipore, #TR-1003-G)
• Puromycin (Invivogen, #ANT-PR-1)
• 6-well and T75 tissue culture-treated assay plates
• Flow cytometer or fluorescence microscope

The lentiviruses were produced from HEK293T cells in 90% DMEM medium containing 10% FBS.

Lentiviruses are shipped on dry ice. For long term storage, it is recommended to store the virus at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Optimizing transduction assays. Easily track transduction efficiency over time. Generation of GFP Reporter stable cell lines

The lentiviruses are produced with the third generation 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.

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.

Cell Signaling Pathway

Cell Line