CAREER: Engineering thermal biosensors for remote control of mammalian cells

使用活细胞作为治疗剂需要严格控制制造过程。严格的控制意味着准确和快速的感知和反应。该项目的目标是设计一种具有温度敏感活性的蛋白质。这种蛋白质可以作为传感器或开关来对局部温度波动做出反应。将这种蛋白质功能与细胞调控网络连接起来，可以让研究人员快速操纵细胞新陈代谢。一种来自真菌的蛋白质将成为模型系统。与当地学区的合作将导致针对高中生的新教育模块。这些活动将促进未被充分代表的群体在STEM教育和职业中的参与。这项提议的中心假设是，一种真菌蛋白质，其活性自然地受温度和光的控制，可以被设计成纯粹对温度敏感，并且这种受温度调节的活动可以被调节并与转录和翻译后过程耦合。具体地说，这项工作将开发一种蛋白质，其从质膜上的解离可以通过温度控制。在目标1中，将测量和模拟工程蛋白膜定位的温度依赖关系。在目标2中，将调整温度敏感度，以便可以在与哺乳动物细胞相关的温度范围(37°C)内控制膜的定位。在目标3中，将开发将膜定位的热控制与控制基因表达或翻译后修饰的细胞内生物化学相结合的分子电路，这将为各种下游应用提供热控制的模块化策略。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Using live cells as therapeutic agents requires strict control of the manufacturing process. Strict control implies accurate and rapid sensing and response. The objective of this project is to engineer a protein with temperature-sensitive activity. The protein could act as a sensor or as a switch to respond to local temperature fluctuations. Connecting this protein function to cellular regulatory networks could allow researchers to rapidly manipulate cell metabolism. A protein from a fungus will be the model system. Collaborating with the local school district will lead to new educational modules directed at high school students. These activities will enhance participation of underrepresented groups in STEM education and careers.The central hypothesis of this proposal is that a fungal protein, whose activity is naturally controlled by both temperature and light, can be engineered to be purely thermo-responsive, and that this thermally-regulated activity can be tuned and coupled to transcriptional and post-translational processes. Specifically, this work will develop a protein whose dissociation from the plasma membrane is controllable by temperature. In Aim 1, the temperature dependence of membrane localization of the engineered protein will be measured and modeled. In Aim 2, the temperature sensitivity will be tuned so that membrane localization can be controlled across temperature ranges relevant for mammalian cells ( 37 °C). In Aim 3, molecular circuits will be developed that couple thermal control of membrane localization to intracellular biochemistry that controls gene expression or posttranslational modifications, which will provide modular strategies for thermal control for a wide variety of downstream applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.