Principal Investigator:
Benjamin Campbell
Background & Unmet Need
- Fluorescent proteins (FPs) are used for a broad range of biology research applications, including as reporters of gene activity and to visualize proteins in live cells
- Most FPs lack the ability to withstand the adverse conditions in the cell that rapidly denature biological structures, including superfolder GFP
- Modern microscopy techniques such as CLEM and ExM, place even greater demands on FPs due to secondary fixation and staining chemicals such as osmium tetroxide that degrade protein functionality
- Most widely used FPs are susceptible to misfolding due to exposure to higher temperatures for a greater lengths of time and photo bleach quickly
- Unmet Need: There is a need for stable FPs that can withstand chaotropic conditions and may be used for various biochemical applications
Technology Overview
- The Technology: Development of extremely stable FPs that withstand chaotropic conditions
- The engineered FPs Hyperfolder YFP (hfYFP) and LSSmGFP successfully survive CLEM sample preparation and ExM techniques
- hfYFP contains no cysteines, is chloride insensitive, and tolerates aldehyde and osmium tetroxide fixation better than common FPs
- The robust nature of hfYFP and LSSmGFP make them ideal FPs for the development of biosensors
- PoC Data: The chemical stability of hfYFP was employed to devise a fluorescence-assisted protein purification strategy, enabling all steps of denaturing affinity chromatography to be visualized using UV or blue light
Technology Applications
- Fluorescence-assisted protein purification
- Live cell imaging and the development of improved biosensors
- Stable FPs for use with CLEM and ExM techniques
Technology Advantages
- Compatible with osmium tetroxide and aldehyde fixation protocols
- Chloride insensitive
- Improved acid resistance
- Greater thermodynamic stability and faster refolding than existing FPs
Publications
Resources
Intellectual Property
Patents
- PCT Application Filed WO2024015362A1: "Chemically stable fluorescent proteins for advanced microscopy"
Cornell Reference
- 10194
Contact Information
For additional information please contact
Jeffrey James
Associate Director, Business Development and Licensing
Phone: (646) 962-4194
Email: jaj268@cornell.edu