Click beetle luciferase mutant and near infrared naphthyl-luciferins for improved bioluminescence imaging
Nature Communications , Volume 9 - Issue 1
The sensitivity of bioluminescence imaging in animals is primarily dependent on the amount of photons emitted by the luciferase enzyme at wavelengths greater than 620 nm where tissue penetration is high. This area of work has been dominated by firefly luciferase and its substrate, D-luciferin, due to the system's peak emission (~ 600 nm), high signal to noise ratio, and generally favorable biodistribution of D-luciferin in mice. Here we report on the development of a codon optimized mutant of click beetle red luciferase that produces substantially more light output than firefly luciferase when the two enzymes are compared in transplanted cells within the skin of black fur mice or in deep brain. The mutant enzyme utilizes two new naphthyl-luciferin substrates to produce near infrared emission (730 nm and 743 nm). The stable luminescence signal and near infrared emission enable unprecedented sensitivity and accuracy for performing deep tissue multispectral tomography in mice.
|This work was funded by the European Commission 7th Framework Programme; grant id fp7/612360 - Molecular Imaging of Brain Pathophysiology (BRAINPATH), This work was funded by the European Commission 7th Framework Programme; grant id h2020/675743 - Image-Guided Surgery (IGS) and Personalised Postoperative Immunotherapy To Improving Cancer Outcome (ISPIC)|
|Organisation||Department of Radiology|
Hall, M.P, Woodroofe, C.C, Wood, M.G, Que, I, van 't Root, M, Ridwan, R.Y, … Mezzanotte, L. (2018). Click beetle luciferase mutant and near infrared naphthyl-luciferins for improved bioluminescence imaging. Nature Communications, 9(1). doi:10.1038/s41467-017-02542-9