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Autonomous Bioluminescent Expression Of The Bacterial Luciferase Gene Cassette (Lux) In A Mammalian Cell Line, Dan M. Close, Stacey S. Patterson, Steven Ripp, Seung J. Baek, John Sanseverino, Gary S. Sayler
Autonomous Bioluminescent Expression Of The Bacterial Luciferase Gene Cassette (Lux) In A Mammalian Cell Line, Dan M. Close, Stacey S. Patterson, Steven Ripp, Seung J. Baek, John Sanseverino, Gary S. Sayler
Steven Ripp
Background
The bacterial luciferase (lux) gene cassette consists of five genes (luxCDABE) whose protein products synergistically generate bioluminescent light signals exclusive of supplementary substrate additions or exogenous manipulations. Historically expressible only in prokaryotes, the lux operon was re-synthesized through a process of multi-bicistronic, codon-optimization to demonstrate for the first time self-directed bioluminescence emission in a mammalian HEK293 cell line in vitro and in vivo.
Methodology/Principal Findings
Autonomous in vitro light production was shown to be 12-fold greater than the observable background associated with untransfected control cells. The availability of reduced riboflavin phosphate (FMNH2) was identified as the limiting bioluminescence substrate …