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Updated: Jan 1, 2026

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

Published on: July 3, 2013

Examining multiple cellular pathways at once using multiplex hextuple luciferase assaying.

Alejandro Sarrion-Perdigones¹, Lyra Chang^2,3, Yezabel Gonzalez¹

¹Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA.

Nature Communications

|December 15, 2019

Summary

This summary is machine-generated.

Researchers developed a new multiplex luciferase assay to simultaneously track six signaling pathways in cells. This advanced method uses distinct substrates and spectral deconvolution for sensitive, cost-effective pathway analysis.

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Last Updated: Jan 1, 2026

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

Published on: July 3, 2013

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

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Published on: June 1, 2014

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay

Published on: September 7, 2008

Area of Science:

Molecular Biology
Biochemistry
Cellular Signaling

Background:

Simultaneous assessment of multiple cellular signaling pathways requires sensitive reporters with broad dynamic ranges.
Genetically encoded luciferases offer sensitivity, versatility, and cost-effectiveness for such assays.

Purpose of the Study:

To expand multiplexing capabilities in post-lysis endpoint luciferase assays from two to six reporter units.
To develop a novel multiplex hextuple luciferase assay for probing pathway fluxes.
To demonstrate the effectiveness and broad applicability of multiplex luciferase assaying.

Main Methods:

Developed a multiplex hextuple luciferase assay by combining distinct substrates and emission spectra deconvolution.
Engineered a single plasmid for solotransfection, delivering all six luciferase reporter units simultaneously.
Probed pathway fluxes through five transcriptional response elements against a control constitutive promoter.

Main Results:

Successfully expanded luciferase multiplexing to six reporter units.
Demonstrated the ability to monitor effects of siRNA, ligand, and chemical compound treatments on target and other cellular pathways.
Validated the effectiveness and adaptiveness of the multiplex luciferase assaying technique.

Conclusions:

The developed multiplex hextuple luciferase assay enables sensitive, simultaneous monitoring of multiple signaling pathways.
Solotransfection using a single plasmid minimizes experimental errors and facilitates reporter unit delivery.
This assay has broad applications across various research fields for studying cellular pathway dynamics.