What question did this study set out to answer?

To engineer a brighter, physically stable bioluminescent enzyme-substrate system with superior biochemical characteristics for mammalian cellular reporter applications.

August 15, 2012ACS Chemical BiologyOpen Access

Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate

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To engineer a brighter, physically stable bioluminescent enzyme-substrate system with superior biochemical characteristics for mammalian cellular reporter applications.
Engineered the 19 kDa luciferase subunit from the deep-sea shrimp Oplophorus gracilirostris through protein structure optimization to generate NanoLuc.
Synthesized and paired NanoLuc with a novel imidazopyrazinone substrate termed furimazine.
Characterized luminescence output, signal duration, specific activity, thermal stability, and cellular expression dynamics in mammalian cells.
NanoLuc with furimazine increased luminescence expression in mammalian cells ~2.5 million-fold compared to the native enzyme precursor.
Exhibited glow-type luminescence with a signal half-life >2 h and specific activity ~150-fold greater than firefly (Photinus pyralis) or Renilla luciferases.
Demonstrated physical stability up to 55 °C and for >15 h in culture medium at 37 °C, showing no evidence of post-translational modifications or subcellular partitioning.

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Implication

Experimental study demonstrates 2.5 million-fold brighter luminescence using engineered shrimp luciferase NanoLuc in mammalian cells, highlighting superior sensitivity for cellular assays.

Cite This Study

Hall et al. (2012) studied this question.

synapsesocial.com/papers/69d71e64236f4746d4563885 https://doi.org/10.1021/cb3002478

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Implication

Experimental study demonstrates 2.5 million-fold brighter luminescence using engineered shrimp luciferase NanoLuc in mammalian cells, highlighting superior sensitivity for cellular assays.

Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate

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Implication

Cite This Study

Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate

Key Points

Discussion

Implication

Cite This Study

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