2005-010 MUTATED RENILLA LUCIFERASE FOR HIGHER LIGHT OUTPUT AND ALTERED STABILITY

A Novel Renilla-Derived Luciferase with Enhanced Activity and Stability

SUMMARY

UCLA researchers in the Department of Molecular and Medical Pharmacology have developed a novel luciferase variant with enhanced stability and activity.

BACKGROUND

Luciferase proteins first derived from firefly are indispensable in biological research. They are used for in vivo imaging, affinity studies by conjugation to antibodies and as reporter proteins in cell culture experiments. The most commonly used luciferase is a small protein derived from Renilla reniformis. Although it is used routinely as a research tool, there are several limitations, as it is unstable in serum, has a low shelf life and a spectral peak of 482 nm that is unsuitable for in vivo imaging.

INNOVATION

UCLA researchers have optimized the Renilla luciferase protein such that it overcomes the current limitations. The optimized protein is stable with 10-fold higher yields. It is active in serum for 180 hours compared to the original protein, which is stable for less than 10 hours. It also has an optimized spectral peak compatible with in vivo imaging. Their optimized variant is thus suitable for assays that were previously incompatible with luciferase.

APPLICATIONS

• In vivo imaging in animal model systems
• In vitro assays of biological samples such as serum
• Reporter for cell culture assays
• Research tool for Bioluminescence Resonance Energy Transfer (BRET)

ADVANTAGES

• 150-fold enhanced stability
• 10-fold higher light emission
• Optimized emission spectra
• Compatible with in vivo imaging

STATE OF DEVELOPMENT

The protein has been tested extensively for stability and performance in mice, cell culture, serum and in reporter assays.

RELATED MATERIALS

• Loening, A. M., Fenn, T. D., Wu, A. M., and Gambhir, S. S. Consensus guided mutagenesis of Renilla luciferase yields enhanced stability and light output, Protein Eng. Des. Sel., 2006.
• Loening, A. M., Wu, A. M., and Gambhir, S. S. Red-shifted Renilla reniformis luciferase variants for imaging in living subjects, Nature Methods, 2007.