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AssigneesCarnegie Institution of Washington, The Board of Trustees of the Leland Stanford Junior University
InventorsCindy AST, Luke M. OLTROGGE, Wolf B. FROMMER
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The present disclosure provides fluorescent polypeptides containing a fusion of a circularly permuted, first fluorescent protein and a second fluorescent protein, the first fluorescent protein containing a first fluorescent moiety and the second fluorescent protein containg a second fluorescent moiety, wherein the second fluorescent protein is contained in the circularly permuted first fluorescent protein to form a cassette that can be inserted into sites within a sensing protein of interest to form novel biosensors; alternatively, the reference domain can be inserted into an existing single-fluorescent protein-based, intensiometric biosensor in order to make a ratiometric biosensor in one cloning step; nucleic acid sequences encoding the fluorescent polypeptides, fluorescent sensors including the fluorescent polypeptides, and methods of making and using same are described herein.
A fluorescent polypeptide comprising a fusion of a circularly permuted, first fluorescent protein as a sensing domain and a second fluorescent protein as a reference domain, the first fluorescent protein comprising a first fluorescent moiety and the second fluorescent protein comprising a second fluorescent moiety, wherein the reference domain is nested within the sequence of the circularly permuted sensing domain, said first and second fluorescent proteins forming a single cassette which can be inserted into sites within a sensing protein of interest to generate a ratiometric biosensor.
A fluorescent sensor comprising a fluorescent polypeptide, said fluorescent polypeptide comprising a circularly permuted first fluorescent protein as a sensing domain and optionally a second fluorescent protein as a reference domain, said second fluorescent protein, when present, being nested within the sequence of the circularly permuted sensing domain so as to form a ratiometric fluorescent sensor.
Dependencies are inferred from the claim text.