Enhancing Viral Transduction in Natural Killer Cells Using a Cationic Polymer

Abstract

Source: Nanbakhsh, A., et al. Dextran Enhances the Lentiviral Transduction Efficiency of Murine and Human Primary NK Cells. J. Vis. Exp. (2018).

This video demonstrates an assay for cationic polymer-mediated viral transduction in natural killer (NK) cells. The cells are incubated with transgenic lentivirus vectors encoding a fluorescent protein. The addition of a cationic polymer enables the virus to attach to the cells and integrate its RNA into the host genome, expressing the fluorescent protein. The fluorescence is quantified to assess the polymer-induced enhancement of transduction.

Protocol

1. Transduction of murine and human primary NK cells with lentivirus Suspend mouse or human primary NK cells in a 24-well plate at 0.5 × 105/mL of medium in the presence of green fluorescent protein (GFP) lentivirus supernatant at 5, 10, and 20 multiplicity of infection (MOI) and in the presence of Pb (8 µg/mL), PS (8 µg/mL) or dextran (8 µg/mL). Centrifuge the plates at 1,000 × g for 60 min. Without decanting the supernatant, c…

Materials

Dextran	Sigma-Aldrich	90-64-91-9
Polybrene (Pb)	Sigma-Aldrich	TR-1003
Protamine sulfate (PS)	Sigma-Aldrich	p3369
RPMI1640	Corning	10-040-CV
Interferon gamma (IFN-γ )	eBioscience	14-7311-85
Propidium lodide staining solution	BD	51-66211E
Lipofectamine 3000	Thermo Fisher	L3000015
Isoflurane	PHOENIX	NDC 57319-559-05
293T cells	ATCC	CRL-3216
T75 flasks	Corning	430641U
ELISA kits	Ebioscience	00-4201-56