Bioluminescence imaging (BLI) is a powerful technique for visualizing small numbers of cells in vivo.  This technology utilizes luciferase enzymes that catalyze light emission via the oxidation of small molecule substrates (luciferins).  Several luciferase-luciferin pairs exist in the natural world, and many have been adapted for tracking cells and other biological targets in whole animals.  Unfortunately, the optimal luciferases for in vivo imaging utilize the same substrate and cannot be used to distinguish multiple cell types in a single subject.  We are expanding the capabilities of BLI by creating luciferase mutants that accept chemically distinct luciferins.  In this work, mutant luciferases were generated using a combination of rational design and random mutagenesis.  The engineered enzymes were also screened against luciferin analogs to identify novel light-emitting pairs.  Our results are being used to guide BLI studies in live animals and develop additional orthogonal probes.

Generating luciferase libraries for multi-component bioluminescence imaging

243rd National Meeting (2012)

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