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Publication Abstract from PubMed

Anthozoa-class red fluorescent proteins (RFPs) are frequently used as biological markers, with far-red (lambdaem approximately 600-700 nm) emitting variants sought for whole-animal imaging because biological tissues are more permeable to light in this range. A barrier to the use of naturally occurring RFP variants as molecular markers is that all are tetrameric, which is not ideal for cell biological applications. Efforts to engineer monomeric RFPs have typically produced dimmer and blue-shifted variants because the chromophore is sensitive to small structural perturbations. In fact, despite much effort, only four native RFPs have been successfully monomerized, leaving the majority of RFP biodiversity untapped in biomarker development. Here we report the generation of monomeric variants of HcRed and mCardinal, both far-red dimers, and describe a comprehensive methodology for the monomerization of red-shifted oligomeric RFPs. Among the resultant variants is mKelly1 (emission maximum, lambdaem = 656 nm), which, along with the recently reported mGarnet2 [Matela G, et al. (2017) Chem Commun (Camb) 53:979-982], forms a class of bright, monomeric, far-red FPs.

Monomerization of far-red fluorescent proteins.,Wannier TM, Gillespie SK, Hutchins N, McIsaac RS, Wu SY, Shen Y, Campbell RE, Brown KS, Mayo SL Proc Natl Acad Sci U S A. 2018 Nov 27;115(48):E11294-E11301. doi:, 10.1073/pnas.1807449115. Epub 2018 Nov 13. PMID:30425172^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.