title: A near-infrared BiFC reporter for in vivo imaging of protein-protein interactions.
abstract: Studies of protein-protein interactions deep in organs and in whole mammals have been hindered by a lack of genetically encoded fluorescent probes in near-infrared region for which mammalian tissues are the most transparent. We have used a near-infrared fluorescent protein iRFP engineered from a bacterial phytochrome as the template to develop an in vivo split fluorescence complementation probe. The domain architecture-based rational design resulted in an iSplit reporter with the spectra optimal for whole-body imaging, high photostability, and high complementation contrast, which compares favorably to that of other available split fluorescent protein-based probes. Successful visualization of interaction of two known protein partners in a living mouse model suggests iSplit as the probe of choice for noninvasive detection of protein-protein interactions in vivo, whereas its fast intracellular degradation enables time-resolved monitoring of repetitive binding events.
authors.affiliation: Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Grigory S
authors.initials: GS
authors.lastname: Filonov
authors.forename: Vladislav V
authors.initials: VV
authors.lastname: Verkhusha
mesh_terms: D000818:Animals; D019020:Directed Molecular Evolution; D005260:Female; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D008856:Microscopy, Fluorescence / Q000379:methods; D061848:Optical Imaging / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051598:Whole Body Imaging
chemical_list: D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins
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