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Reporter Genes

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...

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Related Experiment Video

Updated: Jun 4, 2026

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Published on: August 6, 2014

Imaging real-time gene expression in Mammalian cells with single-transcript resolution.

Amber L Wells¹, John S Condeelis, Robert H Singer

¹Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.

|March 2, 2011

Summary

The MS2 system enables sensitive single-molecule mRNA detection in cells by tagging reporter mRNA with fluorescent coat proteins. This method achieves high signal-to-noise ratios for accurate imaging.

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Last Updated: Jun 4, 2026

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons

Published on: August 6, 2014

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)

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Published on: March 15, 2013

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Published on: February 25, 2017

Area of Science:

Molecular Biology
Cell Biology
Biotechnology

Background:

The MS2 system is crucial for single-molecule detection within cells.
It utilizes a reporter mRNA with MS2 binding sites (MBS) and a fluorescent MS2 coat protein (MCP-xFP).
High affinity binding of MCP-xFP to MBS tags mRNA for visualization.

Purpose of the Study:

To describe a protocol for transfecting COS-7 cells for single-molecule mRNA detection using the MS2 system.
To detail the co-transfection of reporter RNA and MCP-xFP plasmids for simultaneous expression.
To explain how nuclear sequestration of unbound MCP-xFP enhances signal-to-noise ratio.

Main Methods:

Calcium phosphate precipitation for transfecting COS-7 cells.
Co-transfection of reporter mRNA plasmid (e.g., pRSV-Z-24 MBS-β-actin) and MCP-xFP-NLS plasmid (e.g., pPolII-MCP-GFP-NLS).
Utilizing the Delta T Imaging System for fluorescent particle image acquisition.

Main Results:

Successful co-transfection leading to simultaneous expression of reporter mRNA and MCP-xFP-NLS.
Nuclear sequestration of unbound MCP-xFP-NLS, concentrating tagged mRNA in the cytoplasm.
Achieved high signal-to-noise ratio enabling single mRNA molecule detection.

Conclusions:

The described MS2 system protocol provides optimal sensitivity for single-molecule mRNA detection in cells.
Simultaneous expression and targeted localization of fluorescently tagged mRNA facilitate high-resolution imaging.
This method is effective for visualizing and quantifying individual mRNA molecules within a cellular context.