CAREER: Engineering thermal biosensors for remote control of mammalian cells

点击翻译按钮获取中文摘要

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.