Engineering NFAT-Dependent Reporter T Cells

NK92MI cells were obtained from ATCC and maintained as per the instructions provided. JNG (Jurkat cell line engineered with a NFAT-dependent EGFP reporter gene) was kindly provided by Dr. Arthur Weiss (UCSF) and maintained in RPMI-1640 medium supplemented with 10% FBS^{21 (link)}. 293FT cells were obtained from Invitrogen (ThermoFisher Scientific) and cultured as recommended. Primary T cells were isolated and cultured as described earlier^{4 (link)}. Polybrene and coelenterazine were obtained from Sigma and Nano light technology, respectively. Strep-Tactin-APC (IBA-lifescience: Cat# 6-5010-001); Anti-Flag-FITC (Sigma: F4049); Anti-Strep-tagII-FITC (Genscript: A01736); Biotinylated Protein L (Genscript: M0097); Streptavidin-APC conjugate (Molecular probes: SA1005); APC-conjugated anti-MYC antibody (R&D systems: IC3696A); and Polyethylenimine (Polysciences Inc: 23966) were obtained from the indicated sources.

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Gopalakrishnan R., Matta H., Choi S., Natarajan V., Prins R., Gong S., Zenunovic A., Narasappa N., Patel F., Prakash R., Chaudhary V., Sikri V., Chitnis S.D., Kochegarov A., Wang D., Falat M., Kahn M., Keerthipati P.S., Sharma N., Lenka J., Stieben T.M., Braun J., Batra A., Purvis K., Ito K., Lee J.H., Jeronimo A., Zamora H.M., Membreno A., Qiu Q., Peshin S., Namburu L, & Chaudhary P.M. (2019). A novel luciferase-based assay for the detection of Chimeric Antigen Receptors. Scientific Reports, 9, 1957.

Publication 2019

NK92MI cells 293FT cells Polybrene coelenterazine Anti-Flag-FITC Biotinylated Protein L Polyethylenimine

Anti antibody Cell line Cells Coelenterazine Engineered gene Fitc Jurkat cell Light Molecular probes Polybrene Polyethylenimine Protein Strep Streptavidin T cells

Corresponding Organization :

Other organizations : University of Southern California

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Variable analysis

independent variables

Cell lines used: NK92MI, JNG, 293FT, and primary T cells

dependent variables

Not explicitly mentioned

control variables

Culture conditions for each cell line as per instructions and recommendations
Use of Polybrene and coelenterazine
Use of various antibodies and conjugates: Strep-Tactin-APC, Anti-Flag-FITC, Anti-Strep-tagII-FITC, Biotinylated Protein L, Streptavidin-APC conjugate, APC-conjugated anti-MYC antibody
Use of Polyethylenimine

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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