GFP-like fluorescent proteins (FP) have revolutionized cell and molecular biology, however, they cannot be used for whole body and deep brain in vivo imaging due to poor penetration of short-wavelength light through animal tissue. On the other side, there is a challenge in creating a diverse pool of monomeric versions of these proteins with red-shifted spectrum for protein labeling, biosensor development, and multiplexed imaging. Here, we report a monomeric bacterial-phytochrome-based fluorescent protein, called miRFP2, with excitation and emission bands within the highly transparent near-infrared optical interval ranging from 650 to 900nm. miRFP2 can be readily expressed in the widely used model organism, such as C.elegans, Drosophila Melanogaster, Danio rerio, and mice, and does not require an exogenous supply of its chromophore, biliverdin. In addition, miRFP2 is characterized by higher effective fluorescence brightness, signal-to-background ratio, intracellular stability, and photostability than some earlier generations of phytochrome-derived fluorescent probes than expressed in cultured cells and model organisms; it is suitable for super-resolution imaging techniques. It is spectrally compatible with optogenetic tools and can be used for multiplex imaging, providing a new and versatile imaging tool.

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