A platform for targeted, post-translational control of protein levels in mammalian cells

细胞的生理状态由数千种分子成分的集体行为决定，并且可能根据环境因素而发生巨大变化，从而导致非线性行为。尽管系统和合成生物学取得了重大进展，但由于细胞调节和质量控制机制的可塑性和适应性，细胞中的蛋白质功能仍然难以破译。该项目的总体目标是开发一种平台技术来调节细胞中靶蛋白的积累。该技术有望对蛋白质表达提供精确的动态控制，从而提供研究蛋白质功能的工具。将探索一系列互补策略，以实现对蛋白质表达调节的刺激依赖性控制，并利用该技术来研究复杂的生物过程。因此，这项研究有望促进基础科学的进步，推动生物经济的发展，并推动医疗保健的进步。 通过为代表性不足群体的学生提供广泛的教育和培训机会，该项目还有望支持教育和多样性并造福社会。为了克服现有调节蛋白质稳态水平方法的局限性，研究小组使用双功能系统构建了一个翻译后调节蛋白质表达的平台，该双功能系统由用于目标特异性和选择性的分子识别单元和指定目标蛋白质降解速率和机制的降解信号单元组成。拟议研究的目标是设计这项技术，以生成一个通用平台，通过外源或内源刺激控制蛋白质水平。为了实现这一目标，将追求三个研究目标：1）设计蛋白质消耗的刺激响应性降解，2）通过受控的报告基因消耗增强刺激依赖性电路的动态分辨率，3）破译和操纵缺氧信号网络。该研究计划将用于为大休斯顿地区的高中教师和学生提供培训、研究和指导机会，并改善大学生的教学和学习。

Cells physiologic states are dictated by the collective behavior of thousands of molecular components and may vary dramatically depending on environmental factors, resulting in non-linear behaviors. Despite significant advances in systems and synthetic biology, protein function in cells remains hard to decipher due to the plasticity and adaptability of cellular regulatory and quality control mechanisms. The overarching goal of this project is to generate a platform technology to modulate the accumulation of target proteins in cells. This technology is expected to provide exquisite dynamic control over protein expression, thereby providing a tool to study protein function. A series of complementary strategies will be explored to achieve stimulus-dependent control over modulation of protein expression and to deploy this technology to study complex biological processes. As a result, this study is expected to promote progress of fundamental science , advance the bioeconomy, and enable advances in healthcare. By providing broad reaching educational and training opportunities to students belonging to underrepresented groups, this project is also expected to support education and diversity and to benefit society. To overcome the limitations of existing methods for regulating protein steady state levels, the research team built a platform for modulating protein expression post-translationally using a bifunctional system consisting of a molecular recognition unit for target specificity and selectivity, and a degradation-signaling unit that specifies the rate and mechanism of degradation of the target protein. The goal of the proposed research is to engineer this technology to generate a universal platform for controlling protein levels through exogenous or endogenous stimuli. To achieve this goal, three research objectives will be pursued: 1) engineer stimulus-responsive degradation of protein depletion, 2) enhance the dynamic resolution of stimulus-dependent circuits through controlled reporter depletion, and 3) decipher and manipulate the hypoxia signaling network. This research program will be leveraged to provide training, research, and mentoring opportunities for high school teachers and students from the Greater Houston area, and to improve teaching and learning for college students.