Available Technology
Creating a new generation of monomeric flurescent proteins
Technology:
Single-cell screening assay
Markets Addressed
Green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9kDa) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. It is well known that wild-type GFP is a weak dimer that can be made monomeric by introducing a charged residue at the hydrophobic dimer-dimer interface. Many commonly used fluorescent proteins (even monomeric ones) are not truly monomeric and can cause severe mislocalization/clustering artifacts. Furthermore, the brightest, most advanced GFP variant, called superfolder GFP is indeed not monomeric and caused severe localization artifacts.
Scientists at the Paulsson lab created controlled mutations in specific sites and were able to convert supferfolder GFP into a monomeric tag and hence create a fluorescent protein which had minimal amount of mislocalization or clustering artifacts around it.
Based on the proof of concept with the super-fold GFP, which is, to date, the most advanced GFP currently on the market, the Paulsson lab is currently developing a single cell screening assay that uses a large collection of monomeric mutations. This will enable the identification of specific mutations that may cause artifacts in certain dyes due to low binding selectivity.
Locating those mutations will enable the creation of a new generation of more selective dyes that would have lower rates of mislocalization/clustering artifacts.
Innovations and Advantages
This is a relatively easy single-cell screening assay that can identify monomeric mutations in different florescent dyes targeting different wavelength ranges.
This screening tool, which is based on the oligomerization tendencies of fluorescent proteins, will enable the identification and production of new and improved generations of monomeric fluorescent dyes. It can enable the creation of dyes that would produce less mislocalization/clustering artifacts. And as a result would produce more accurate spatiotemporal results.
Additional Information
Intellectual Property Status: Patent(s) pending
Tweet
Inventor(s):
Landgraf, Dirk
Paulsson, Johan
Categories:
For further information, please contact:
Michal Preminger, Director of Business Development
(617) 432-0920
Reference Harvard Case #4493