Regulated Protein Degradation

项目摘要 这项研究计划的广泛、长期的目标是开发通用的方法来 在蛋白质分子水平上有条件地调节蛋白质的功能，而不是通过靶向 编码目标蛋白质的DNA或信使核糖核酸前体。这些实验方法是高度有效的。 针对目标蛋白质，并提供快速和可调节的控制蛋白质功能使用细胞- 可渗透的小分子。他们的目标是设计出一种叫做破坏稳定的小蛋白结构域 在哺乳动物细胞中表达时迅速降解的结构域。不稳定的不稳定性 结构域被忠实地授予与这些小结构域融合的伙伴蛋白，使研究人员能够 可以预测地控制任何感兴趣的蛋白质的水平。这项研究计划的一个具体目标是 提供新的不稳定结构域，这些结构域源自人类蛋白质，并受FDA- 批准的药物。这项研究的第二个目标是开发一种新的方法来破坏结构域的稳定 调节人体细胞分泌治疗有用的细胞因子和激素。第三个目标 这项研究的重点是开发一种新的方法来调节受 只有当整个融合蛋白是 由它的同源配体稳定。第四个目标建立在生物物理学的基础上， 不稳定结构域开发一类新型的细胞内秒基因编码生物传感器 信使分析物。目前缺乏安全有效的方法来调控基因的表达和 基于基因的疗法的生物活性严重限制了疾病的数量和类型 医生和科学家可以考虑使用细胞和基因疗法进行靶向治疗。新法规 安全有效的不稳定域等方法将极大地扩大 可以通过细胞和基因疗法进行靶向治疗的疾病的宇宙，从而打开 治疗用现有方法无法解决的人类疾病的新领域。

Project Summary The broad, long-term objective of this research program is to develop general methods to conditionally regulate protein function at the level of the protein molecules rather than by targeting the DNA or mRNA precursors that encode a protein-of-interest. These experimental methods are highly specific for the targeted proteins and provide rapid and tunable control of protein function using cell- permeable small molecules. The goal is to engineer small protein domains called destabilizing domains that are rapidly degraded when expressed in mammalian cells. The instability of destabilizing domains is faithfully conferred to partner proteins fused to these small domains, allowing researchers to predictably control the levels of any protein-of-interest. One specific aim of this research program will provide new destabilizing domains that are derived from human proteins and regulated by FDA- approved drugs. A second aim of this research is to develop a new method for destabilizing domains to regulate the secretion of therapeutically useful cytokines and hormones from human cells. A third aim of this research is the development of a new method for the partner proteins regulated by the destabilizing domains to be liberated from the regulatory domain only when the entire fusion protein is stabilized by its cognate ligand. The fourth aim builds upon the biophysics underpinning the destabilizing domains to develop a new class of genetically encoded biosensors for intracellular second messenger analytes. The current lack of safe and effective methods to regulate the expression and biological activity of gene-based therapeutics seriously limits the number and types of diseases that physicians and scientists can contemplate targeting using cell and gene therapy. New regulation methods such as the destabilizing domains that are safe and effective will dramatically expand the universe of diseases that can be targeted for treatment through cell and gene therapy, thus opening new frontiers for treating human diseases that cannot be addressed using existing methods.