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  • Title: Rational design of photoconvertible and biphotochromic fluorescent proteins for advanced microscopy applications.
    Author: Adam V, Moeyaert B, David CC, Mizuno H, Lelimousin M, Dedecker P, Ando R, Miyawaki A, Michiels J, Engelborghs Y, Hofkens J.
    Journal: Chem Biol; 2011 Oct 28; 18(10):1241-51. PubMed ID: 22035793.
    Abstract:
    Advanced fluorescence imaging, including subdiffraction microscopy, relies on fluorophores with controllable emission properties. Chief among these fluorophores are the photoactivatable fluorescent proteins capable of reversible on/off photoswitching or irreversible green-to-red photoconversion. IrisFP was recently reported as the first fluorescent protein combining these two types of phototransformations. The introduction of this protein resulted in new applications such as super-resolution pulse-chase imaging. However, the spectroscopic properties of IrisFP are far from being optimal and its tetrameric organization complicates its use as a fusion tag. Here, we demonstrate how four-state optical highlighting can be rationally introduced into photoconvertible fluorescent proteins and develop and characterize a new set of such enhanced optical highlighters derived from mEosFP and Dendra2. We present in particular NijiFP, a promising new fluorescent protein with photoconvertible and biphotochromic properties that make it ideal for advanced fluorescence-based imaging applications.
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