Red fluorescent protein

Red fluorescent protein (RFP) is a fluorophore that fluoresces red-orange when excited. Several variants have been developed using directed mutagenesis.[1] The original was isolated from Discosoma, and named DsRed. Others are now available that fluoresce orange, red, and far-red.[2]

Red fluorescent protein drFP583
Identifiers
OrganismDiscosoma sp.
Symbol?
UniProtQ9U6Y8

RFP is approximately 25.9 kDa. The excitation maximum is 558 nm, and the emission maximum is 583 nm.[3]

The first fluorescent protein to be discovered, green fluorescent protein (GFP), has been adapted to identify and develop fluorescent markers in other colors. Variants such as yellow fluorescent protein (YFP) and cyan fluorescent protein (CFP) were discovered in Anthozoa.[4]

Issues with fluorescent proteins include the length of time between protein synthesis and expression of fluorescence. DsRed has an maturation time of around 24 hours,[1] which can make it unusable for many experiments that take place in a shorter time frame. Additionally, DsRed exists in a tetrameric form, which can affect the function of proteins to which it is attached. Genetic engineering has improved the utility of RFP by increasing the speed of fluorescent development and creating monomeric variants.[3][4] Improved variants of RFP include mFruits (mCherry, mOrange, mRaspberry), mKO, TagRFP, mKate, mRuby, FusionRed, mScarlet and DsRed-Express.[4][5]

DsRed has been shown to be more suitable for optical imaging approaches than EGFP.[6]

References
  1. Bevis, Brooke J.; Glick, Benjamin S. (2002). "Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed)". Nature Biotechnology. 20 (1): 83–87. doi:10.1038/nbt0102-83. ISSN 1546-1696. PMID 11753367.
  2. Miyawaki, Atsushi; Shcherbakova, Daria M; Verkhusha, Vladislav V (October 2012). "Red fluorescent proteins: chromophore formation and cellular applications". Current Opinion in Structural Biology. 22 (5): 679–688. doi:10.1016/j.sbi.2012.09.002. ISSN 0959-440X. PMC 3737244. PMID 23000031.
  3. Remington, S. James (1 January 2002). "Negotiating the speed bumps to fluorescence". Nature Biotechnology. 20 (1): 28–29. doi:10.1038/nbt0102-28. PMID 11753356.
  4. Piatkevich, Kiryl D.; Verkhusha, Vladislav V. (2011). "Guide to Red Fluorescent Proteins and Biosensors for Flow Cytometry". Methods in Cell Biology. 102: 431–461. doi:10.1016/B978-0-12-374912-3.00017-1. ISBN 9780123749123. ISSN 0091-679X. PMC 3987785. PMID 21704849.
  5. Bindels, Daphne S; Haarbosch, Lindsay (2017). "mScarlet: a bright monomeric red fluorescent protein for cellular imaging". Nature Methods. 14 (1): 53–56. doi:10.1038/nmeth.4074. ISSN 1548-7105. PMID 27869816.
  6. Böhm I, Gehrke S, Kleb B, Hungerbühler M, Müller R, Klose KJ, Alfke H (2019). "Monitoring of tumor burden in vivo by optical imaging in a xenograft SCID mouse model: evaluation of two fluorescent proteins of the GFP-superfamily". Acta Radiol. 60 (3): 315–326. doi:10.1177/0284185118780896. PMID 29890843.
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