In Vivo Imaging of Far-red Fluorescent Proteins after DNA Electrotransfer to Muscle Tissue
Author Information
Author(s): Hojman Pernille, Eriksen Jens, Gehl Julie
Primary Institution: Centre of Inflammation and Metabolism, Department of Infectious Diseases, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark
Hypothesis
We want to characterize the novel far-red fluorescent protein Katushka as a marker for gene expression using time domain fluorescence in vivo imaging.
Conclusion
Katushka offers excellent evaluation of transfection efficacy with a signal much brighter than GFP, but is not suitable for long-term detection of in vivo transgenic expression.
Supporting Evidence
- Katushka expression peaked 1 week after transfection and returned to background levels within 4 weeks.
- Katushka was 3-fold brighter than GFP at 1 week after DNA electrotransfer.
- Electrotransfer of 5 μg of Katushka plasmid resulted in a large increase in in vivo fluorescent intensity.
Takeaway
The study tested a new bright protein called Katushka to see how well it shows gene activity in muscles, and found it works well at first but doesn't last long.
Methodology
DNA electrotransfer was used to deliver plasmids encoding Katushka and GFP into muscle tissue, followed by in vivo imaging to assess expression.
Limitations
Katushka expression was not as stable as GFP expression, which limited its long-term utility.
Participant Demographics
7–9-week-old female C57Black/6 or NMRI mice.
Digital Object Identifier (DOI)
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