Optically Modulated Fluorescent Proteins

描述(由申请人提供)：分子特异性荧光成像的进步彻底改变了对细胞中生化过程的跟踪。然而，一个持续存在的问题是，在存在来自内源或外源荧光团的背景荧光时，信噪比相对较低。我们建议采用一种称为光调制的信号处理技术来提高这一比率。我们将设计和合成基于蓝色荧光蛋白的蛋白质，这种蛋白质将快速转化为绿色吸收形式。这种形式可以通过长波长照射转换回荧光形式，产生可用于光学调制和锁定放大的跟踪信号。通过将已知的蓝色荧光蛋白或可光切换蛋白突变为正确的吸收/异构化特征，我们将开发出能够真正进行细胞内单分子成像的光学可调制荧光蛋白。与公共卫生的相关性：公共卫生声明光学调节荧光蛋白荧光蛋白彻底改变了对细胞中生化过程的跟踪，2008年诺贝尔奖认可了这一点，科学文献中有100,000多篇关于使用荧光蛋白的参考文献。利用从已知荧光蛋白中提取的新蛋白质，我们建议开发一组新的蛋白质，将它们与一种称为光学调制的信号处理技术相耦合，以极大地提高灵敏度。在近分子水平上跟踪细胞通路动态的能力将使依赖于细胞运输缺陷的疾病状态取得重大进展，包括癌症和自身免疫性疾病。

DESCRIPTION (provided by applicant): Advances in molecularly specific fluorescent imaging have revolutionized the tracking of biochemical processes in cells. A continuing problem, however, has been the relatively poor signal-to-noise ratio in the presence of background fluorescence from endogenous or exogenous fluorophores. We propose to employ a signal processing technique called optical modulation to improve that ratio. We will design and synthesize proteins based upon a blue fluorescent protein which will undergo fast conversion to a green-absorbing form. This form can be converted back to the fluorescent form by long- wavelength irradiation, producing a tracking signal which can be used for optical modulation and lock-in amplification. By mutating known blue fluorescent proteins or photoswitchable proteins toward the right absorption/isomerizations characteristics, we will develop optically modulatable fluorescent proteins capable of true intracellular single molecule imaging. PUBLIC HEALTH RELEVANCE: Public health statement Optically Modulated Fluorescent Proteins Fluorescent proteins have revolutionized the tracking of biochemical processes in cells, as recognized by the 2008 Nobel Prize in the field and by the presence of over 100,000 references to the use of fluorescent proteins in the scientific literature. Using new proteins derived from known fluorescent proteins, we propose to develop a new set of proteins, coupling them to a signal processing technique called optical modulation, to greatly improve sensitivity. The ability to follow the dynamics of cellular pathways at the near molecular level will allow significant progress to be made in disease states that are dependent upon defects in cell transport, including cancer and autoimmune diseases.