Directed
Molecular Evolution Reveals Gaussia Luciferase
Variants with Enhanced Light Output Stability
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Abstract
Gaussia
Luciferase (Gluc) has proven to be a powerful mammalian
cell reporter for monitoring numerous biological processes in immunology,
virology, oncology, and neuroscience. Current limitations of Gluc
as a reporter include its emission of blue light, which is absorbed
by mammalian tissues, limiting its use in vivo, and a flash-type bioluminescence
reaction, making it unsuited for high-throughput applications. To
overcome these limitations, a library of Gluc variants was generated
using directed molecular evolution and screened for relative light
output, a shift in emission spectrum, and glow-type light emission
kinetics. Several variants with a 10–15 nm shift in their light
emission peak were found. Further, a Gluc variant that catalyzes a
glow-type bioluminescence reaction, suited for high-throughput applications,
was also identified. These results indicate that molecular evolution
could be used to modulate Gluc bioluminescence reaction characteristics