분자영상 및 방사화학

  • [Cancer Res] Fluorophore-NanoLuc BRET Reporters Enable Sensitive In Vivo Optical Imaging and Flow Cytometry for Monitoring Tumorigenesis

    University of North Carolina at Chapel Hill / Antonio L. Amelio*

  • 출처
    Cancer Res
  • 등재일
    2015 Dec 1
  • 저널이슈번호
    75(23):5023-33. doi: 10.1158/0008-5472.CAN-14-3538. Epub 2015 Sep 30.
  • 내용

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    Abstract


    Fluorescent proteins are widely used to study molecular and cellular events, yet this traditionally relies on delivery of excitation light, which can trigger autofluorescence, photoxicity, and photobleaching, impairing their use in vivo. Accordingly, chemiluminescent light sources such as those generated by luciferases have emerged, as they do not require excitation light. However, current luciferase reporters lack the brightness needed to visualize events in deep tissues. We report the creation of chimeric eGFP-NanoLuc (GpNLuc) and LSSmOrange-NanoLuc (OgNLuc) fusion reporter proteins coined LumiFluors, which combine the benefits of eGFP or LSSmOrange fluorescent proteins with the bright, glow-type bioluminescent light generated by an enhanced small luciferase subunit (NanoLuc) of the deep-sea shrimp Oplophorus gracilirostris. The intramolecular bioluminescence resonance energy transfer that occurs between NanoLuc and the fused fluorophore generates the brightest bioluminescent signal known to date, including improved intensity, sensitivity, and durable spectral properties, thereby dramatically reducing image acquisition times and permitting highly sensitive in vivo imaging. Notably, the self-illuminating and bifunctional nature of these LumiFluor reporters enables greatly improved spatiotemporal monitoring of very small numbers of tumor cells via in vivo optical imaging and also allows the isolation and analyses of single cells by flow cytometry. Thus, LumiFluor reporters are inexpensive, robust, noninvasive tools that allow for markedly improved in vivo optical imaging of tumorigenic processes. Cancer Res; 75(23); 5023-33.  

     

     

    Author information

    Schaub FX1, Reza MS2, Flaveny CA3, Li W1, Musicant AM4, Hoxha S5, Guo M2, Cleveland JL1, Amelio AL6.

    1 Department of Tumor Biology, Moffitt Cancer Center and Research Institute, Tampa, Florida.

    2 Department of Cancer Biology, The Scripps Research Institute, Scripps Florida, Jupiter, Florida.

    3 Department of Pharmacological & Physiological Science, School of Medicine, Saint Louis University, St. Louis, Missouri.

    4 UNC Biological and Biomedical Sciences Graduate Program, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.

    5 Scripps Graduate Program, The Scripps Research Institute, Scripps Florida, Jupiter, Florida.

    6 Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina. Biomedical Research Imaging Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina. antonio_amelio@unc.edu.

     

  • 편집위원

    Bioluminescence resonance energy transfer (BRET) 기반의 reporter를 이용한 in vivo optical 영상법으로, ‘NanoLuc’을 이용한 새로운 형광영상기법을 소개하였습니다.

    2016-01-05 15:06:17

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