Imaging G-Quadruplexes in Live Cells
Author Information
Author(s): Bradford Tigerlily, Summers Peter A., Majid Aatikah, Sherin Petr S., Lam Jeff Yui Long, Aggarwal Savyasanchi, Vannier Jean-Baptiste, Vilar Ramon, Kuimova Marina K.
Primary Institution: Molecular Sciences Research Hub, Department of Chemistry, Imperial College London
Hypothesis
Thioflavin T and Thiazole Orange will display distinct fluorescence lifetimes when binding to different DNA topologies.
Conclusion
Thioflavin T is a promising probe for detecting G-Quadruplex DNA and RNA in live cells using fluorescence lifetime imaging microscopy.
Supporting Evidence
- Thioflavin T showed a significant increase in fluorescence lifetime when binding to G4 DNA compared to duplex DNA.
- Thiazole Orange displayed a 600- to 850-fold enhancement in fluorescence upon binding to G4 DNA.
- The study demonstrated that ThT is suitable for G4 detection in live cells due to its stable fluorescence lifetime.
Takeaway
This study shows that special dyes can help scientists see important structures in DNA while the cells are still alive.
Methodology
The study used fluorescence lifetime imaging microscopy (FLIM) to visualize G-Quadruplexes in live U2OS cells with Thioflavin T and Thiazole Orange.
Limitations
TO showed poor reproducibility in live cells due to aggregation, making it unsuitable for G4 detection.
Participant Demographics
U2OS human bone osteosarcoma epithelial cells were used.
Statistical Information
P-Value
p < 0.001
Confidence Interval
1.71–2.10 ns
Statistical Significance
p < 0.001
Digital Object Identifier (DOI)
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