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• W4313391912 startingPage "677" @default.
• W4313391912 abstract "Bioluminescence (BL) is an excellent optical readout for bioassays and molecular imaging. Herein, we accomplished new near infrared bioluminescence resonance energy transfer (NIR-BRET) templates for monitoring molecular events in cells with higher sensitivity. We first identified the best resonance energy donor for the NIR-BRET templates through the characterization of many coelenterazine (CTZ)-marine luciferase combinations. As a result, we found that NLuc-DBlueC and ALuc47-nCTZ combinations showed luminescence in the blue emission wavelength with excellent BL intensity and stability, for example, the NLuc-DBlueC and ALuc47-nCTZ combinations were 17-fold and 22-fold brighter than their second highest combinations, respectively, and were stably bright in living mammalian cells for at least 10 min. To harness the excellent BL properties to the NIR-BRET systems, NLuc and ALuc47 were genetically fused to fluorescent proteins (FPs), allowing large blue-to-red shifts, such as LSSmChe, LSSmKate2, and LSSmNep (where LSS means Large Stokes Shift). The excellent LSSmNep-NLuc combination showed approximately 170 nm large resonance energy shift from blue to red. The established templates were further utilized in the development of new NIR-BRET systems for imaging steroid hormone activities by sandwiching the ligand-binding domain of a nuclear receptor (NR-LBD) between the luciferase and the FP of the template. The NIR-BRET systems showed a specific luminescence signal upon exposure to steroid hormones, such as androgen, estrogen, and cortisol. The present NIR-BRET templates are important additions for utilizing their advantageous imaging of various molecular events with high efficiency and brightness in physiological samples." @default.
• W4313391912 created "2023-01-06" @default.
• W4313391912 creator A5018880989 @default.
• W4313391912 creator A5033559819 @default.
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• W4313391912 date "2022-12-30" @default.
• W4313391912 modified "2023-09-26" @default.
• W4313391912 title "Near-Infrared Imaging of Steroid Hormone Activities Using Bright BRET Templates" @default.
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• W4313391912 doi "https://doi.org/10.3390/ijms24010677" @default.
• W4313391912 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/36614119" @default.
• W4313391912 hasPublicationYear "2022" @default.
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