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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.

Tagging and Fusion Proteins

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

Labeling DNA Probes

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Related Experiment Video

Updated: Apr 20, 2026

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

One-step CRISPR-based Strategy for Endogenous Gene Tagging in Drosophila melanogaster

Published on: January 26, 2024

Endogenous gene tagging with fluorescent proteins.

John Fetter¹, Andrey Samsonov, Nathan Zenser

¹Cell-Based Assays/Reporter Cell Lines, Sigma-Aldrich Research Biotech, 2909 Laclede Avenue, Saint Louis, MO, 63103, USA.

Methods in Molecular Biology (Clifton, N.J.)

|November 20, 2014

Summary

This summary is machine-generated.

Genome editing enables precise monitoring of disease genes by inserting reporter tags. This preserves natural gene expression for accurate live-cell analysis and drug discovery.

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Area of Science:

Genomics
Molecular Biology
Biotechnology

Background:

Understanding disease mechanisms often requires studying gene function within cells.
Traditional methods like overexpression or random integration can alter native gene expression and regulation.
Accurate monitoring of endogenous gene activity is crucial for disease research and drug development.

Purpose of the Study:

To highlight the utility of genome editing for creating precise cellular models.
To emphasize the advantages of reporter gene knock-ins for studying endogenous gene expression.
To showcase the potential of these engineered cells for drug discovery.

Main Methods:

Utilizing modern genome editing tools to insert reporter sequences into specific genomic locations.
Creating fusion proteins expressed at physiological levels under endogenous regulatory control.
Developing cell-based assays for monitoring gene activity in live cells.

Main Results:

Knock-in cells maintain native gene expression and regulation, unlike transient transfection or random integration methods.
Fusion proteins accurately reflect endogenous protein expression profiles.
Live-cell detection of target activity is possible, avoiding fixation artifacts.

Conclusions:

Genome editing provides a superior method for creating cellular models to study disease gene mechanisms.
Reporter-tagged endogenous genes allow for precise, real-time monitoring of cellular processes.
Engineered cell lines facilitate effective compound screening for novel drug discovery.