What is a reporter gene and how is a reporter gene used?

Reporter genes are genes whose products can be readily assayed subsequent to transfection, and can be used as markers for screening successfully transfected cells, for studying regulation of gene expression, or serve as controls for standardizing transfection efficiencies.

How are reporter genes used in vitro promoter assay?

The main purpose of the reporter gene assay is to investigate the promoter of a gene of interest, i.e. the regulation of its expression. This can be done by linking the promoter of interest to an easily detectable gene, such as the gene for firefly luciferase, which catalyses a reaction that produces light.

How does luciferase reporter gene work?

By transfection, a DNA construct with the gene’s promoter and a coding region of the luciferase reporter gene enters the cells. When this protein activates transcription, the cell will produce luciferase enzyme. After the addition of a lysis buffer and a substrate, a luminometer quantifies the luciferase activity.

How do you perform a reporter assay on a gene?

To perform the reporter assay, you clone the regulatory region of your gene-of-interest (X) upstream of the luciferase gene in one of these expression vectors, introduce that resulting vector DNA into cells, and let the cells grow for a period of time.

How are reporter genes introduced into an organism?

To introduce a reporter gene into an organism, scientists place the reporter gene and the gene of interest in the same DNA construct to be inserted into the cell or organism. For bacteria or prokaryotic cells in culture, this is usually in the form of a circular DNA molecule called a plasmid.

How does luciferase reporter gene enter the cell?

By transfection, a DNA construct with the gene’s promoter and a coding region of the luciferase reporter gene enters the cells. Another DNA construct introduced into the cells consists of a coding region of the protein of interest.

How are expression levels of reporter genes measured?

The reporter gene construct contains one or more gene regulatory elements to be analyzed, the sequence for the reporter gene, and the sequences required for the transcription of functional mRNA. Upon introduction of the reporter construct into cells, expression levels of the reporter gene are monitored through a direct assay of