Nat. Biotechnol.
34, 760-767.
A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo.
2016
Chu, J., Oh, Y., Sens, A., Ataie, N., Dana, H., Macklin, J.J., Laviv, T., Welf, E.S., Dean, K.M., Zhang, F., Kim, B.B., Tang, C.T., Hu, M., Baird, M.A., Davidson, M.W., Kay, M.A., Fiolka, R., Yasuda, R., Kim, D.S., Ng, H.L. and Lin, M.Z.
Notes: The authors characterize a novel orange-red fluorescent protein (CyOFP1), which serves as an efficient acceptor for resonance energy transfer from NanoLuc® luciferase. An optimized fusion protein of CyOFP1 and NanoLuc® (Antares) was demonstrated to function as a highly sensitive bioluminescent reporter in vivo, producing substantially brighter signals from deep tissues than firefly luciferase and other bioluminescent proteins. During fluorometry, 50,000 cells in 10 μL Live Cell Imaging Solution were loaded into glass capillary tubes and gently pelleted by centrifugation for 15 s at 500g. 10 μL of Live Cell Imaging Solution with 20 μM D-luciferin, coelenterazine, ViviRen, or furimazine was then added. For BRET6 and Antares-expressing mice, tail vein injections were performed with 333 nmol (123 μg) of furimazine in 120 μL of 8% glycerol, 12% ethanol, 10% hydroxypropyl-β-cyclodextrin, and 35% polyethylene glycol 400 in water. (4760)
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