Analysis and Bioengineering of Optogenetic Rhodopsins

光遗传学视紫红质的分析和生物工程

• 批准号: 1264434
• 负责人: Kenneth Rothschild
• 金额: $ 59.8万
• 依托单位: Trustees of Boston University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1264434&HistoricalAwards=false
• 关键词: Analysis; Bioengineering; Optogenetic; Rhodopsins

1264434RothschildThe research will involve three laboratories at the BU Photonics Center (Rothschild, Erramilli and Han) and utilize advanced spectroscopic and biophysical/bioengineering techniques along with collaborators at UC Davis Center for Biophotonics, MIT, Harvard University, and University of Texas. A central feature of the research will be the use of time-resolved FTIR difference spectroscopic, ultra-fast laser spectroscopy and confocal near-IR Raman techniques to measure molecular changes in optogenetic rhodopsins upon light excitation on a time-scale of seconds to femtoseconds. Preliminary work has demonstrated that a variety of optogenetic rhodopsins can be reconstituted into proteolipid membranes including nanolipidparticles and studied using advanced spectroscopic methods to obtain detailed molecular information about their function. In addition, spectroscopic studies have been extended to living cells allowing these proteins to be studied in their native environment in order to determine molecule effects of such factors as membrane potential. Collaborators on this project include: Dr. Ed Boyden at MIT, widely recognized as one of the co-founders of optogenetics and a leading developer of new neurophotonic technology; Dr. Matt Coleman at UCD, who has developed methods of expressing rhodopsins into nanolipidparticles (NLPs) and forming microcrystals suitable for coherent x-ray diffraction imaging (CXDI), Dr. John Spudich, at the U. Texas Med. School, a pioneer in the field of microbial rhodopsins and co-discoverer of channelrhodopsins and Dr. Adam Cohen, whose laboratory at Harvard University has recently demonstrated that rhodopsins can be used to monitor transmembrane voltage in living nerves.

1264434Rothschild 该研究将涉及 BU 光子学中心的三个实验室（Rothschild、Erramilli 和 Han），并与加州大学戴维斯分校生物光子学中心、麻省理工学院、哈佛大学和德克萨斯大学的合作者一起利用先进的光谱和生物物理/生物工程技术。该研究的一个核心特点是使用时间分辨傅立叶变换红外光谱、超快激光光谱和共焦近红外拉曼技术来测量光遗传学视紫红质在几秒到飞秒时间尺度的光激发下的分子变化。初步工作表明，多种光遗传学视紫红质可以重构为包括纳米脂质颗粒在内的蛋白脂质膜，并使用先进的光谱方法进行研究以获得有关其功能的详细分子信息。此外，光谱研究已扩展到活细胞，允许在其天然环境中研究这些蛋白质，以确定膜电位等因素的分子效应。该项目的合作者包括： 麻省理工学院的 Ed Boyden 博士，被广泛认为是光遗传学的联合创始人之一和新神经光子技术的领先开发商；都柏林大学的马特·科尔曼 (Matt Coleman) 博士开发了将视紫红质表达为纳米脂质颗粒 (NLP) 并形成适合相干 X 射线衍射成像 (CXDI) 的微晶体的方法，而德克萨斯大学医学院的约翰·斯普迪奇 (John Spudich) 博士则开发了这种方法。 School 是微生物视紫红质领域的先驱和通道视紫红质的共同发现者，他和哈佛大学的 Adam Cohen 博士的实验室最近证明视紫红质可用于监测活体神经的跨膜电压。