Enhancing Viral Transduction in Natural Killer Cells Using a Cationic Polymer
Transcripción
Take natural killer, or NK, cells. Add lentivirus vectors comprising viral enzymes and engineered RNA molecules that encode a fluorescent protein.
Add cationic polymers and sediment the cells and viruses.
The polymers neutralize the negative charges on both the virus and the cell, facilitating membrane fusion.
The RNA is reverse-transcribed into DNA and integrated into the host genome, enabling fluorescent protein expression.
Add media supplemented with the cytokine interleukin-2, or IL-2.
IL-2 triggers intracellular signaling that promotes NK cell survival and proliferation while preserving transgene expression.
Transfer the mixture to a well coated with antibodies that bind to specific immunoreceptors on NK cells, triggering them to release antiviral cytokines.
Quantify the released cytokines to assess the polymer's impact on cytokine production.
Add fluorescent dyes to label the DNA of necrotic cells.
Using flow cytometry, identify live cells and quantify the fluorescent protein's signal to measure the efficiency of polymer-induced transduction.
