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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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Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

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Improved RNA toehold switch sensitivity using the NanoLuc complementation reporter.

Katherinne J Diaz¹, Julian Jarquin¹, Aleksandr Petrosyan¹

¹Biology Department, California State University, Northridge, Northridge, California, United States.

Micropublication Biology

|January 22, 2025

Summary

RNA toehold switches act as biosensors detecting specific RNA triggers. Using the NanoBiT reporter enhances sensitivity compared to sfGFP for broad RNA detection applications.

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Area of Science:

Molecular Biology
Synthetic Biology
Biotechnology

Background:

RNA toehold switches are programmable genetic devices that regulate gene expression in response to specific RNA sequences.
These switches are utilized in cell-free systems for biosensing applications, including the detection of viral and bacterial RNA.
Reporter proteins like sfGFP are commonly used to signal the activation of toehold switches.

Purpose of the Study:

To evaluate the NanoLuc complementation reporter system, NanoBiT, as a more sensitive alternative to sfGFP for RNA toehold switch applications.
To demonstrate the broad applicability of NanoBiT in enhancing RNA biosensor sensitivity.

Main Methods:

The study involved designing and implementing RNA toehold switches in a cell-free expression system.
NanoBiT and sfGFP were used as reporter systems to quantify toehold switch activation.
The sensitivity of NanoBiT was compared against sfGFP in response to specific RNA triggers.

Main Results:

NanoBiT demonstrated significantly improved sensitivity in detecting RNA triggers compared to sfGFP.
The NanoBiT reporter system proved effective across various RNA toehold switch designs.
This enhanced sensitivity allows for more robust and reliable RNA detection.

Conclusions:

The NanoBiT reporter system offers superior sensitivity for RNA toehold switch-based biosensors.
NanoBiT represents a valuable advancement for cell-free biosensing and diagnostics.
This technology has broad potential for detecting diverse RNA targets with increased precision.