NSF/MCB-BSF: Development of novel CRAC channel photoswitches

NSF/MCB-BSF：开发新型 CRAC 通道光电开关

• 批准号: 2116412
• 负责人: Michael Kienzler
• 金额: $ 79.62万
• 依托单位: University of Connecticut
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2116412&HistoricalAwards=false
• 关键词: NSF; MCB; BSF; Development; novel

This project will provide new molecular tools and understanding about Calcium Release Activated Calcium (CRAC) channels. These are key proteins that affect calcium entry and signaling in cells. Calcium is one of the most important signaling molecules in living cells. Understanding calcium entry and signaling has important implications in understanding numerous cellular processes including migration, proliferation, and gene expression. This NSF-BSF project is led by researchers at the University of Maine in the US, in collaboration with researchers at the Technion in Haifa, Israel. As part of this international collaboration, graduate and undergraduate students will be trained in interdisciplinary research at the interface of organic chemistry, molecular biology, and biophysics. Interactions and training will be facilitated through regular video meetings and travel to the collaborator’s laboratory in Israel. As a complimentary goal, the Broader Impacts of this proposal will increase research participation of traditionally underrepresented groups in STEM at the University of Maine, particularly women and first-generation college students.The overarching goal of this project is to better understand and control the function of CRAC channels, which are an essential component of store-operated calcium entry (SOCE), an evolutionary conserved mechanism that regulates intracellular calcium levels in metazoans. The first aim of this proposal will combine synthetic chemistry and molecular biology in order to develop small molecule Light Operated CRAC Channel Inhibitors (LOCIs), which will provide rapid and reversible control of the function of both native and genetically engineered CRAC channels. A set of LOCIs with different photophysical properties will be synthesized and evaluated using electrophysiological and live cell imaging techniques. As there are multiple CRAC channel isoforms, the second Aim of this project will combine synthetic chemistry and genetic engineering to develop new types of LOCIs with two-component optogenetic constructs that will enable covalent tethering of LOCIs to the cell membrane or directly to different CRAC channel components. These constructs will enable the control of CRAC channel activity in a cell-type and isoform specific manner and with high spatial and temporal precision. Finally, LOCIs will be used as light activated modulators of calcium dependent biological processes, particularly cell migration and gene expression.This collaborative US/Israel project is supported by the US National Science Foundation and the Israeli Binational Science Foundation.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.

该项目将提供新的分子工具，并了解钙释放活化钙（CRAC）通道。这些是影响细胞中钙进入和信号传导的关键蛋白。钙是活细胞中最重要的信号分子之一。了解钙进入和信号传导对理解包括迁移，增殖和基因表达在内的众多细胞过程具有重要意义。该NSF-BSF项目由美国缅因州大学的研究人员与以色列海法技术的研究人员合作。作为这项国际合作的一部分，研究生和本科生将接受有机化学，分子生物学和生物物理学界面的跨学科研究培训。互动和培训将通过定期的视频会议准备，并前往以色列合作者的实验室。 As a complimentary goal, the Broader Impacts of this proposal will increase research participation of traditionally underrepresented groups in STEM at the University of Maine, particularly women and first-generation college students.The overarching goal of this project is to better understand and control the function of CRAC channels, which are an essential component of store-operated calcium entry (SOCE), an evolutionary conserved mechanism that regulates intracellular calcium levels in metazoans.该提案的第一个目的是结合合成化学和分子生物学，以开发小分子光操作的CRAC通道抑制剂（LOCIS），该抑制剂（LOCIS）将对天然和遗传性CRAC通道的功能快速，可逆地控制。将使用电生理和活细胞成像技术合成和评估具有不同光物理特性的一组LOCI。由于有多种CRAC通道同工型，该项目的第二个目的将结合合成化学和基因工程，以开发新型的LOCIS和两组分组的光遗传学构建体，这些构造将使LOCIS能够将LOCIS的连接到细胞膜或直接到不同的CRAC通道成分。这些构建体将以细胞类型和同工型的特定方式以及具有高空间和临时精度来控制CRAC通道活性。最后，LOCIS将用作依赖钙依赖生物学过程的光激活调节剂，尤其是细胞迁移和基因表达。该协作的美国/以色列项目得到了美国国家科学基金会和以色列双原则科学基金会的支持。该奖项反映了NSF的法定任务，并通过评估了基金会的智力效果，并诚实地对其进行了评估。