Advantages
- The developed method offers highly sensitive in vivo imaging of viral infection dynamics.
- The manipulating Akaluc luciferase gene for the same codon-usages is a trick of generation of the recombinant virus carrying the reporter gene.
Background and Technology
Analysis of the time course of viral infection dynamics in vivo is one of the techniques that is useful for elucidating the pathology, tissue tropism, or efficacy of antiviral drugs. It is possible to analyze the dynamics in vivo (in vivo imaging) by artificially generating a reporter virus in which a fluorescent protein (reporter) gene is recombined with a viral gene and infecting a model animal.
The fluorescent protein NanoLuc and general luciferases have short emission wavelengths, so they cannot detect light deep inside the body and cannot be used for in vivo imaging of viral infection dynamics. Akaluc luciferase has an emission wavelength of 670-680 nm, so it can be used for in vivo imaging.
We generated a reporter virus by recombining the AkaLuc gene with the SARS-CoV-2 viral genome. However, sufficient viral proliferation ability was not obtained, so we performed codon optimization to match the GC content of the viral genome and the GC content of the recombinant Akaluc gene. As a result, we were able to generate a reporter virus with high proliferation ability and succeeded in in vivo imaging of the infection dynamics of SARS-CoV-2.
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Patent and Reference
- Patent pending (unpublished yet)
- Tamura et al., iScience. (2024) 29;27(5):109647
Principal Investigator
Takasuke FUKUHARA (Graduate School of Medicine, Hokkaido University)
Current Stage and Next Step
- We generated the Akaluc reporter virus using SARS-CoV-2 variants, infected hamsters, and successfully performed in vivo imaging of the infection dynamics. Furthermore, using a mouse model, we confirmed the infection suppression effect of the mRNA vaccine by in vivo
- We plan to generate codon-optimized Akaluc reporter viruses for other viruses and continue research using in vivo
- We are currently seeking antiviral drug and vaccine development companies that wish to conduct collaborative research using this reporter virus and in vivo imaging technology.
Project.BK-04948