A luciferase reporter assay is a cell-based, enzymatic experiment that utilizes an ectopically expressed luciferase in cultured cells or in live animals, with the presence of its substrate luciferin, to generate luminescence in response to gene regulation at the level of transcription. This assay can be easily formatted for high-throughput sample analysis. The reagents are commercially available and routinely used in various applications. We have automated a luciferase reporter assay on the Opentrons OT-2 platform to measure activation of NF-kB in HeLa cells. The Python script-based protocols were developed to perform cell seeding, reporter construct transfection, and enzyme-catalytic reaction to produce detectable signals for quantification with desirable quality of sample handling and minimal hands-on time.
Keyphrases
- high throughput
- single cell
- cell cycle arrest
- induced apoptosis
- crispr cas
- signaling pathway
- pi k akt
- oxidative stress
- lps induced
- cell death
- nuclear factor
- deep learning
- endothelial cells
- transcription factor
- cell proliferation
- mesenchymal stem cells
- nitric oxide
- inflammatory response
- immune response
- structural basis