Bioluminescence imaging (BLI) is among the most dynamic imaging modalities for visualizing whole cells and gene expression patterns in vivo. This technique captures light emission from the luciferase-catalyzed oxidation of small molecule luciferins with sensitive cameras. While powerful, current options for multiplexed BLI in preclinical models are limited by the number of native luciferase/luciferin pairs optimized for in vivo use. Our lab aims to expand the bioluminescent toolkit by pairing mutant firefly luciferases with synthetic luciferin analogs, to biochemically resolve multiple targets via sequential imaging. Several generations of mutant luciferase libraries were screened against sterically and electronically modified families of luciferins, in order to find orthogonal pairs. Promising luciferases were expressed recombinantly for biochemical analysis with their respective luciferins. These pairs were evaluated for their potential for multicomponent in vivo imaging using protein and tissue culture models.

Mutant firefly luciferases catalyze light emission with complementary synthetically modified luciferins

249th National Meeting (2015)

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