Abstract
The bacterial luciferase gene cassette (lux) is unique among bioluminescent bioreporter systems due to its ability to synthesize and/or scavenge all of the substrate compounds required for its production of light. As a result, the lux system has the unique ability to autonomously produce a luminescent signal, either continuously or in response to the presence of a specific trigger, across a wide array of organismal hosts. While originally employed extensively as a bacterial bioreporter system for the detection of specific chemical signals in environmental samples, the use of lux as a bioreporter technology has continuously expanded over the last 30 years to include expression in eukaryotic cells such as Saccharomyces cerevisiae and even human cell lines as well. Under these conditions, the lux system has been developed for use as a biomedical detection tool for toxicity screening and visualization of tumors in small animal models. As the technologies for lux signal detection continue to improve, it is poised to become one of the first fully implantable detection systems for intra-organismal optical detection through direct marriage to an implantable photon-detecting digital chip. This review presents the basic biochemical background that allows the lux system to continuously autobioluminesce and highlights the important milestones in the use of lux-based bioreporters as they have evolved from chemical detection platforms in prokaryotic bacteria to rodent-based tumorigenesis study targets. In addition, the future of lux imaging using integrated circuit microluminometry to image directly within a living host in real-time will be introduced and its role in the development of dose/response therapeutic systems will be highlighted.
Highlights
Bacterial bioluminescence, commonly known as the lux reaction, is the most widely distributed luminescent mechanism on the planet [1] and, this process of bacterial light production has been observed for centuries, it was not until the mid 1900s that it began to be evaluated scientifically [2,3]
This review will highlight the development of the lux cassette from a curiosity observed in marine bacteria, through its extensive use as a bacterial bioreporter system and modification for expression in eukaryotic organisms, up to its recent demonstration as the only fully autonomous, substrate-free bioluminescent reporter system available in the eukaryotic host background
While the use of bacterial bioluminescence as a reporter system has been employed for quite a long time, it is still a continually developing reporter system
Summary
Commonly known as the lux reaction, is the most widely distributed luminescent mechanism on the planet [1] and, this process of bacterial light production has been observed for centuries, it was not until the mid 1900s that it began to be evaluated scientifically [2,3]. While initially expression of the bacterial genes was unsuccessful, through rearrangement of the lux cassette gene expression pattern and improvement of expression efficiency via codon-optimization and the addition of specialized linker regions, these hurdles were overcome and the lux reaction was demonstrated to occur in the lower eukaryote Saccharomyces cerevisiae [5] Building upon this early success of eukaryotic expression, the luxAB genes were further engineered to express in a human cell line, leading to the emergence of the lux system as a truly multifunctional reporter system similar to the more commonly employed firefly luciferase system [6]. This review will highlight the development of the lux cassette from a curiosity observed in marine bacteria, through its extensive use as a bacterial bioreporter system and modification for expression in eukaryotic organisms, up to its recent demonstration as the only fully autonomous, substrate-free bioluminescent reporter system available in the eukaryotic host background. This feedback sysstem servess to regulatte biolumin nescent producction in thesse organism ms
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