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Implementing VIPER for Imaging Cellular Proteins by Fluorescence Microscopy
Julia K Doh1, Caroline A Enns2, Kimberly E Beatty1,3
1Department of Biomedical Engineering, Oregon Health & Science University, Portland, Oregon 97239, USA.
Bio-Protocol
|July 16, 2020
Summary
New genetically-encoded Versatile Interacting Peptide (VIP) tags, like VIPER, enable specific protein labeling for multicolor cellular imaging in live and fixed cells. This protocol details VIPER implementation for receptor and intracellular target visualization.
Area of Science:
- Cell Biology
- Molecular Imaging
- Biotechnology
Background:
- Genetically-encoded tags are essential for advanced cellular imaging techniques.
- Versatile Interacting Peptide (VIP) tags offer novel capabilities for multicolor and multi-scale imaging.
- Existing methods may have limitations in specificity or application scope.
Purpose of the Study:
- To provide a detailed protocol for implementing VIPER technology in cellular imaging.
- To demonstrate the application of VIPER for visualizing live-cell receptors and fixed-cell intracellular targets.
- To establish VIPER as a versatile tool for protein labeling in diverse imaging contexts.
Main Methods:
- Utilizing VIPER, a genetically-encoded tag based on coiled-coil heterodimerization.
- Implementing a two-peptide system: one as the genetic tag, the other delivering a reporter.
- Applying the protocol for live-cell imaging of receptors and fixed-cell imaging of intracellular targets.
Main Results:
- Demonstrated rapid and specific heteromer formation for selective protein labeling.
- Successfully visualized receptors on live cells using VIPER technology.
- Effectively imaged intracellular targets in fixed cells with VIPER.
Conclusions:
- VIPER technology provides a robust and specific method for genetically-encoded protein labeling.
- The protocol facilitates multicolor and multi-scale imaging applications in both live and fixed cells.
- VIPER represents a significant advancement in tools for cellular and molecular imaging.
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