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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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Updated: Jul 26, 2026

High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay

High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay

Published on: June 1, 2014

A dual-luciferase reporter system for studying recoding signals

G Grentzmann¹, J A Ingram, P J Kelly

¹Howard Hughes Medical Institute, University of Utah, Salt Lake City 84112, USA.

RNA (New York, N.Y.)

|June 18, 1998

Summary

A novel dual-luciferase reporter system enables efficient measurement of translation recoding mechanisms. This assay accurately quantifies frameshifting and readthrough efficiencies in vitro and in vivo.

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Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

Translation coupling efficiency is crucial for gene expression regulation.
Recoding mechanisms like frameshifting and readthrough modulate protein synthesis.
Existing methods for measuring these efficiencies can be complex or lack internal controls.

Purpose of the Study:

To develop a novel reporter system for measuring translation coupling efficiency.
To establish a convenient and reliable assay for frameshifting and readthrough.
To enable direct comparison of recoding efficiencies in vitro and in vivo.

Main Methods:

A dual-luciferase reporter construct was engineered, placing a recoding sequence between Renilla and Firefly luciferase genes.
Luciferase activities were measured in the same tube, allowing for normalization.
The system was applied to study recoding in HIV-1 gag-pol, MMTV gag-pro, MuLV gag-pol, and human antizyme.

Main Results:

The normalized ratio of Renilla to Firefly luciferase activity directly correlates with recoding efficiency.
This dual-luciferase assay provides a reliable and convenient method for measuring translation coupling.
The system is suitable for both in vitro and in vivo applications, facilitating direct comparisons.
The assay demonstrated applicability to various biological systems, including viral and human genes.

Conclusions:

The developed dual-luciferase reporter system is the first enzymatic assay suitable for in vitro translation coupling efficiency measurements.
This assay offers a robust platform for high-throughput screening of translation signals.
It allows for an intimate comparison of in vivo and in vitro recoding mechanisms from a single construct.