CD16A/NFAT Reporter Lentivirus (ADCC Assay)

*Transduction units (TU), based on the number of drug resistant or fluorescent HEK293FT cells after transduction with the virus.

Vector Layout

The CD16A/NFAT Immunotherapy Reporter Lentivirus provides a convenient tool for researchers to perform ADCC Assays and create custom cell lines using their cells of interest. The reporter system functions by detecting NFAT-mediated transcriptional activity in response to Fc engagement of CD16A expressed on the cell surface. The all-in-one reporter construct allows the user to easily generate stable reporter cell lines that constitutively express a cell surface receptor/ligand of interest (CD16A) and drug selection marker (Puro or BSD), and conditionally express a fluorescent (GFP or RFP) and secreted Gaussia luciferase (GLuc) dual reporter in response to transcriptional activation of the specific signaling pathway (NFAT). Constructs with constitutively expressed fluorescent proteins instead of a drug selection marker are also available for users who prefer to isolate transduced cells by FACS. This product is supplied as high-titer lentiviral particles (third generation), which have been purified by PEG precipitation and sucrose gradient centrifugation. The lentiviral particles are VSV-G pseudotyped and are ideal for difficult-to-transfect cells including primary or thawed cells.

Product Data

Figure 1. Jurkat cells (5×10^5) were transduced with 100 μl of the CD16A(V158)-BSD/NFAT-GFP-GLuc lentiviral particles (SKU: TRV-0002-6S) in a 12-well plate with 6 μg/ml polybrene, followed by centrifugation for 2 hours. The cells were then incubated for an additional 2-4 hours. The entire culture was transferred to a T25 flask and cultured in 10 ml RPMI1640 medium for 3 days. The medium was then changed, and 10 μg/ml blasticidin was added to select for drug-resistant ADCC-Jurkat cells. These cells were co-cultured with target Raji cells in the presence of 100 ng/ml of anti-CD20 antibody (BioXCell SKU: SIM0008) or an isotype control antibody for 16 hours before flow cytometry analysis. 

Figure 2. ADCC Assay CD16/NFAT Jurkat Cell Line (EGFP/GLuc Dual-Reporter) cells were co-cultured with target Raji cells in the presence of 100 ng/ml of anti-CD20 antibody (BioXCell SKU: SIM0008) or an isotype control antibody for 16 hours followed by luminescence detection on the Spectramax iD3 with the LipExoGen coelenterazine solution.