Reconstructing Promoter Activity From Lux Bioluminescent Reporters

The bacterial Lux system is used as a gene expression reporter. It is fast, sensitive and non-destructive, enabling high frequency measurements. Originally developed for bacterial cells, it has been adapted for eukaryotic cells, and can be used for whole cell biosensors, or in real time with live an...

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Bibliographic Details
Published inbioRxiv
Main Authors Iqbal, Mudassar, Doherty, Neil, Page, Anna M L, Qazi, Saara N A, Ajmera, Ishan, Lund, Peter A, Kypraios, Theodore, Scott, David J, Hill, Philip J, Stekel, Dov J
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 15.03.2017
Subjects
Bioluminescence
Biosensors
Gene expression
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Summary:The bacterial Lux system is used as a gene expression reporter. It is fast, sensitive and non-destructive, enabling high frequency measurements. Originally developed for bacterial cells, it has been adapted for eukaryotic cells, and can be used for whole cell biosensors, or in real time with live animals without the need for slaughter. However, correct interpretation of bioluminescent data is limited: the bioluminescence is different from gene expression because of nonlinear molecular and enzyme dynamics of the Lux system. We have developed a modelling approach that, for the first time, allows users of Lux assays to infer gene transcription levels from the light output. We show examples where a decrease in bioluminescence would be better interpreted as a switching off of the promoter, or where an increase in bioluminescence would be better interpreted as a longer period of gene expression. This approach could benefit all users of Lux technology.
DOI:10.1101/117093