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Bioluminescent Bacterial Imaging In Vivo

Bioluminescent Bacterial Imaging In Vivo

Published on: November 4, 2012

An improved pathway for autonomous bioluminescence imaging in eukaryotes.

Ekaterina S Shakhova^1,2, Tatiana A Karataeva^1,2, Nadezhda M Markina^1,2

¹Planta LLC, Moscow, Russia.

|January 22, 2024

Summary

Researchers optimized a fungal bioluminescence pathway for brighter light in plants, fungi, and animals. This breakthrough enables enhanced imaging capabilities in various host organisms.

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Area of Science:

Biochemistry
Molecular Biology
Synthetic Biology

Background:

The bioluminescence pathway from Neonothopanus nambi fungus allows for self-sustained light emission in engineered eukaryotes.
Native fungal enzymes in heterologous hosts exhibit limited performance, restricting luminescence brightness.

Purpose of the Study:

To engineer optimized versions of the fungal bioluminescence pathway.
To significantly enhance bioluminescence intensity in diverse eukaryotic hosts.
To enable advanced imaging applications, including longitudinal video-rate imaging.

Main Methods:

Enzyme engineering and directed evolution of key components of the Neonothopanus nambi bioluminescence pathway.
Heterologous expression and characterization of optimized pathways in plant, fungal, and mammalian cell systems.
Assessment of luminescence output and imaging capabilities using advanced microscopy techniques.

Main Results:

Optimized pathway variants demonstrated a one-to-two order of magnitude increase in bioluminescence brightness compared to native enzymes.
Sustained and bright luminescence was achieved across plant, fungal, and mammalian host systems.
The enhanced bioluminescence enabled high-resolution, longitudinal video-rate imaging of biological processes.

Conclusions:

Engineered bioluminescence pathways offer a substantial improvement in light output for various eukaryotic hosts.
This advancement provides a powerful tool for non-invasive, real-time biological imaging and monitoring.
The optimized system has broad applications in research, diagnostics, and biotechnology.