In vivo bioluminescence imaging of natural bacteria within deep tissues via ATP-binding cassette sugar transporter

Most existing bioluminescence imaging methods can only visualize the location of engineered bacteria in vivo, generally precluding the imaging of natural bacteria. Herein, we leverage bacteria-specific ATP-binding cassette sugar transporters to internalize luciferase and luciferin by hitchhiking them on the unique carbon source of bacteria. Typically, the synthesized bioluminescent probes are made of glucose polymer (GP), luciferase, Cy5 and ICG-modified silicon nanoparticles and their substrates are made of GP and D-luciferin-modified silicon nanoparticles. Compared with bacteria with mutations in transporters, which hardly internalize the probes in vitro (i.e., ~2% of uptake rate), various bacteria could robustly engulf the probes with a high uptake rate of around 50%. Notably, the developed strategy enables ex vivo bioluminescence imaging of human vitreous containing ten species of pathogens collected from patients with bacterial endophthalmitis. By using this platform, we further differentiate bacterial and non-bacterial nephritis and colitis in mice, while their chemiluminescent counterparts are unable to distinguish them.

 

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This passage describes a new bioluminescence imaging method that leverages bacteria-specific ATP-binding cassette sugar transporters to internalize luciferase and luciferin, allowing for the visualization of natural bacteria in vivo. The synthesized bioluminescent probes are made of glucose polymer (GP), luciferase, Cy5 and ICG-modified silicon nanoparticles and their substrates are made of GP and D-luciferin-modified silicon nanoparticles. The strategy was found to enable ex vivo bioluminescence imaging of human vitreous containing ten species of pathogens collected from patients with bacterial endophthalmitis. The platform was also able to differentiate bacterial and non-bacterial nephritis and colitis in mice, while their chemiluminescent counterparts were unable to distinguish them. The uptake rate of the probes was around 50% in various bacteria, compared to bacteria with mutations in transporters, which hardly internalize the probes in vitro (i.e., ~2% of uptake rate).

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S6580	D-Luciferin Potassium Salt	D-luciferin is the natural substrate of the enzyme luciferase (Luc) which catalyzes the production of the typical yellow-green light of fireflies. The 560 nm chemiluminescence from this reaction peaks within seconds, with light output that is proportional to luciferase concentration when the substrate luciferin is present in excess. The luciferase (luc) gene is a popular reporter gene for research and agent screening. Chemiluminescent techniques are virtually background-free, making the luc reporter gene ideal for detecting low-level gene expression. In addition to its role as a reporter of gene expression, luciferase is commonly used in an extremely sensitive assay for ATP.

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