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Significance Although cyanobacteriochrome photoreceptors are potentially useful molecules for bioimaging and optogenetics because of their compactness and spectral diversity, most cyanobacteriochromes have a drawback to need a chromophore unavailable in the mammalian cells. To overcome this significant drawback, this study focused on a mammalian intrinsic chromophore, biliverdin. We succeeded in conversion of biliverdin-rejective cyanobacteriochromes into biliverdin acceptable molecules by replacement of only four residues. Furthermore, molecular basis for the biliverdin incorporation has been revealed by the X-ray crystallography. One such engineered molecule was shown to function as a near-infrared fluorescent probe for in vivo imaging in living mice. This study provides a rational strategy to obtain molecules applicable to bioimaging and optogenetics.

Rational conversion of chromophore selectivity of cyanobacteriochromes to accept mammalian intrinsic biliverdin