Enhancing Viral Transduction in Natural Killer Cells Using a Cationic Polymer
Published: April 30, 2024
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…
Divulgazioni
The authors have nothing to disclose.
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 |
Tags
Citazione di questo articolo
Enhancing Viral Transduction in Natural Killer Cells Using a Cationic Polymer. J. Vis. Exp. (Pending Publication), e22223, doi: (2024).