Optically Modulated Fluorescent Proteins

描述（由申请人提供）：分子特异性荧光成像的进展彻底改变了细胞中生化过程的跟踪。然而，一个持续存在的问题是，在存在来自内源性或外源性荧光团的背景荧光的情况下，信噪比相对较差。我们建议采用一种称为光调制的信号处理技术来提高该比率。我们将设计和合成基于蓝色荧光蛋白的蛋白质，该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。通过突变已知的蓝色荧光蛋白或光开关蛋白朝向正确的吸收/异构化特性，我们将开发能够真正的细胞内单分子成像的光学可调荧光蛋白。公共卫生相关性：荧光蛋白质彻底改变了细胞中生化过程的跟踪，这一点得到了该领域2008年诺贝尔奖的认可，并且在科学文献中有超过100，000篇关于荧光蛋白质使用的参考文献。使用来自已知荧光蛋白的新蛋白质，我们建议开发一组新的蛋白质，将它们与称为光调制的信号处理技术相结合，以大大提高灵敏度。在近分子水平上跟踪细胞途径动力学的能力将允许在依赖于细胞转运缺陷的疾病状态中取得重大进展，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.