Real Time Monitoring of Signaling Pathways in Biological Cells
Abstract
The goal of this research project was to determine the efficacy of EviTags as in situ nanoprobes. The experimental design used a fluorescence resonance energy transfer (FRET)-based approach to show how EviTags can monitor real-time cellular events, in particular, cell surface receptor trafficking and mRNA stability. EviTags were hypothesized to be excellent FRET donors when paired with organic dyes as acceptors because of their unique optical properties. The broad absorption spectra of EviTags allows for low wavelength excitation that does not overlap with organic dye excitation and consequently eliminates acceptor signal bleed through contamination. EviTags also have narrow full width-half max emission spectra with minimal, if any, spectral overlap with acceptor emission spectra thereby eliminating donor signal bleed through contamination. To evaluate EviTags as probes for cell surface receptor trafficking, the transferrin receptor was used as a model system. To address how EviTags can be used to measure mRNA stability, molecular beacon technology was employed. The results of the first phase of this project show EviTags to be highly efficient FRET donors, capable of being delivered into cells, and to be promising in situ nanoprobes.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 14, 2005
- Accession Number
- ADA430353
Entities
People
- Brian D. Cohen
- Louise J. Brogan