Bioluminescence resonance energy transfer (BRET) systems and methods of use thereof
AssigneeStanford University
InventorsSanjiv S. Gambhir, Abhijit De
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Abstract
Briefly described, embodiments of this disclosure include bioluminescence resonance energy transfer (BRET) systems, methods of detecting a protein-protein interaction, noninvasive methods for detecting the interaction of a first protein with a second protein within a living animal, methods to determine the efficacy of a test compound administered to modulate the interaction of a first protein with a second protein in a living animal, BRET vectors, kits relating to each of the above, transgenic cell or progeny thereof and/or animals relating to each of the above, and the like.
Claims6
A bioluminescence resonance energy transfer (BRET) system comprising:
- a first fusion protein including a first target protein and a bioluminescence donor molecule, wherein the bioluminescence donor molecule is a mutated Renilla Luciferase protein having the polypeptide sequence SEQ ID NO:3;
- a second fusion protein including a second target protein and a fluorescent acceptor molecule;
- and a bioluminescence initiating compound, wherein the bioluminescence initiating compound is a coelenterazine.
A bioluminescence resonance energy transfer (BRET) system comprising:
- a first fusion protein including a first target protein and a bioluminescence donor molecule, wherein the bioluminescence donor molecule is a mutated Renilla Luciferase protein selected from:
- SEQ ID NO:3 or conservatively modified variants of SEQ ID NO:3 containing the specific mutations A55T, C124A, S130A, K136R, A143M, M185V, M253L and S287L, wherein the conservatively modified variant of SEQ ID NO:3 emits bioluminescence having a wavelength between about 360 to 460 nm when contacted with coelenterazine, and has an enhanced bioluminescence level relative to the wild-type Renilla reniformis luciferase protein when contacted with the bioluminescence compound, coelenterazine;
- a second fusion protein including a second target protein and a fluorescent acceptor molecule;
- and a bioluminescence initiating compound, wherein the bioluminescence initiating compound is a coelenterazine.
Dependencies are inferred from the claim text.