Coelenterazine-Utilizing Luciferase Reporters with Extremely Stable Glow-Type Bioluminescent Signal
Field: Protein engineering
Challenge:
Luminescence reporters are useful and popular tools in research and biotechnologies, allowing easy-to-use, rapid, safe, and sensitive monitoring of biological, biochemical, and chemical processes by measuring an optical signal. The luminescence process is initiated by the excitation of luminescent molecules to a high-energy state. Return to the ground state with lower energy is accompanied by the release of a photon, observable as light emission.
Bioluminescence is a fascinating phenomenon involving the emission of light by a living creature. There is enormous interest in harnessing bioluminescent systems to design ultrasensitive optical bioassays.
Despite many advances and improvements in luciferases, two major limitations are preserved. First, the flash-type of the bioluminescence signal with a short half-life prevents prolonged signal collection and analyses. Second, moderate substrate affinity does not allow analyses at lower substrate concentrations with the maintained level of the detected bioluminescence signal.
Tech overview:
The present invention relates to modified protein sequences of coelenterazine-utilizing Renilla-type luciferases utilizing widely accessible, non-cytotoxic, and low-cost coelenterazine luciferin. These novel luciferases exhibit extremely stable glow-type light emission, high substrate affinity, low product inhibition factor, and modulated emission maximum. The luciferases are designed to provide ultrasensitive and stable bioluminescent signals in a wide range of laboratory applications and bioassay formats.
Benefits:
- extremely stable glow-type bioluminescent signal
- high affinity towards luciferin substrate
- low product inhibition
- enhanced thermostability
- high resistance to inactivation
- high expressibility & solubility
- no cytotoxicity
- tunable light emission enabling multiplexing
- highly compliant to crystalize & high diffraction quality
Applications:
- wide range of biosensor and reporter applications and bioassay formats
- in vitro and in vivo bioimaging applications
- long-term bioimaging with stable bioluminescent signal
- suitable for bioluminescence resonance energy transfer (BRET) technologies
- codon-optimized gene reporters for both bacterial and mammalian expression
- his-tagged luciferases enabling affinity purification & immunodetection
- recombinantly-produced and purified luciferases ready for use
Commercial opportunity:
- The University is looking for a commercial partner who would develop the technology into a commercial product – as a kit for in vitro and in vivo bioimaging.
- The technology is also available for licensing.
IP protection status:
PCT application filed