title: Directed molecular evolution reveals Gaussia luciferase variants with enhanced light output stability.
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.
authors: Experimental Therapeutics and Molecular Imaging Laboratory, Neuroscience Center, Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, United States.
authors: M Hannah
authors: MH
authors: Degeling
authors: M Sarah S
authors: MS
authors: Bovenberg
authors: Grant K
authors: GK
authors: Lewandrowski
authors: Mark C
authors: MC
authors: de Gooijer
authors: Carmen L A
authors: CL
authors: Vleggeert-Lankamp
authors: Marie
authors: M
authors: Tannous
authors: Casey A
authors: CA
authors: Maguire
authors: Bakhos A
authors: BA
authors: Tannous
mesh_terms: D000595:Amino Acid Sequence; D019020:Directed Molecular Evolution; D008027:Light; D008156:Luciferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009154:Mutation
chemical_list: D008156:Luciferases