Mutagenesis study of a Bacteriophytochrome – insights for the development of labels for optical imaging

2026 · doi:10.64898/2026.03.03.708993
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Abstract Bacteriophytochromes (BphPs) find increasing interest as near-infrared (NIR) labels for imaging. Applications range from whole animal imaging to cell-to super-resolution microscopy. Here we present a comprehensive study of a BphP from Rhizobium etli (ReBphP) allowing mutant-based insights into BphP photophysics. This is complemented by QM/MM-optimized deep-learning structure predictions of ReBphP and variants in their photoswitched states, rationalizing the effects of variants. Pertaining to imaging, based on our study we identify a bright and far red-shifted BphP for imaging in mammalian cells as well as a fluorescent photoswitching BphP. Utilizing the latter, we introduce photoswitching fluorescence background suppression for in vivo whole animal fluorescence imaging achieving higher contrast over background than possible utilizing non-switching labels. Competing Interest Statement VN is a founder and equity owner of Maurus OY, sThesis GmbH, Spear UG, Biosense Innovations P.C. and I3 Inc. All other authors declare no competing interests

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last seen: 2026-05-20T01:45:00.602351+00:00