Due to its immunogenicity in mice and humans the Fluc system does not currently fulfil its true potential in biomedical research. Most BLI studies lack biological relevance and therefore have reduced statistical power, which could be dramatically increased by reporter engineering of the Fluc system. We - and others have heavily engineered the Fluc system for improved in vivo imaging properties. Our infra-luciferin system is unique and helps overcome problems with the validity of BLI in biomedical research by providing more accurate near-infrared cell imaging.
However, to date, there has been no attempt to engineer Flucs that better shielded from murine immune system. Since this aim would lead to a reduction in animal use it would fit well with the ‘imaging bioengineered tissues’ theme within the NC3Rs and in-line with the aims of Cardiff University. This work would impact on the NC3Rs in two ways: a. by improving the uptake of, and extending the use of BLI technologies in stem cell research, and b. by improving the biological relevance and informativeness of stem cell research, hence improving the link between preclinical testing and clinical application of stem cell therapy. Improved translation will lead impact on prevention, diagnosis and treatment strategies in humans and lead to refinement and reduction in the use of small and large animals in wide-ranging research, from the scale of individual experiments to larger scale scientific planning and longer term directions in biomedical and biological sciences.
Hardinge P and Murray JAH. (2019). Reduced False Positives and Improved Reporting of Loop-Mediated Isothermal Amplification using Quenched Fluorescent Primers. Scientific Reports 9:7400. doi: 10.1038/s41598-019-43817-z
Halliwell LM et al. (2018). ΔFlucs: Brighter Photinus pyralis firefly luciferases identified by surveying consecutive single amino acid deletion mutations in a thermostable variant. Biotechnology and Bioengineering 115(1):50-59. doi: 10.1002/bit.26451
Anderson JC et al. (2017). Convergent synthesis and optical properties of near-infrared emitting bioluminescent infra-luciferins. RSC advances 7(7):3975-82. doi: 10.1039/c6ra19541e
- Engagement Activities: Pint of Science 2018