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Updated: Feb 2, 2026
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Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
Published on: January 6, 2015
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Monomerization of far-red fluorescent proteins
Timothy M Wannier1, Sarah K Gillespie2, Nicholas Hutchins2
1Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125; timothy_wannier@hms.harvard.edu steve@mayo.caltech.edu.
Summary
Researchers developed bright, monomeric red fluorescent proteins (RFPs) for improved biological imaging. These new far-red variants overcome limitations of existing tetrameric RFPs, enhancing their utility in cell biology and whole-animal studies.
Area of Science:
- Biochemistry
- Molecular Biology
- Biophotonics
Background:
- Red fluorescent proteins (RFPs) from Anthozoa are valuable biological markers.
- Far-red emitting RFPs are desirable for whole-animal imaging due to tissue light penetration.
- Oligomeric structures of native RFPs hinder their application in cell biology.
Purpose of the Study:
- To engineer monomeric variants of far-red dimeric RFPs (HcRed and mCardinal).
- To establish a methodology for monomerizing red-shifted oligomeric RFPs.
- To expand the toolkit of bright, monomeric, far-red fluorescent proteins for research.
Main Methods:
- Monomerization of HcRed and mCardinal using protein engineering.
- Development of a comprehensive protocol for monomerizing oligomeric RFPs.
- Characterization of spectral properties and brightness of engineered variants.
Main Results:
- Successful generation of monomeric variants of HcRed and mCardinal.
- Introduction of mKelly1, a bright monomeric RFP with an emission maximum at 656 nm.
- Demonstration of a versatile methodology applicable to various red-shifted oligomeric RFPs.
Conclusions:
- Engineered monomeric RFPs offer significant advantages over their oligomeric counterparts.
- mKelly1 and mGarnet2 represent a new class of bright, monomeric, far-red fluorescent proteins.
- The developed methodology facilitates the untapped potential of RFP biodiversity for biomarker development.
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