Analysis and Bioengineering of Optogenetic Rhodopsins

1264434 Rothschild这项研究将涉及波士顿大学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。这项研究的一个主要特点将是使用时间分辨FTIR差分光谱、超快激光光谱和共焦近红外拉曼技术，在几秒到飞秒的时间尺度上测量光激发时光遗传视紫红素的分子变化。初步工作表明，各种光遗传视紫红质可以重组成包括纳米脂颗粒在内的蛋白脂膜，并使用先进的光谱方法进行研究，以获得关于其功能的详细分子信息。此外，光谱研究已经扩展到活细胞，允许在它们的自然环境中研究这些蛋白质，以便确定诸如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，他被广泛认为是光遗传学的联合创始人之一，也是新的神经光子技术的领先开发者；UCD的Matt Coleman博士，他开发了将视紫红质表达成纳米脂质颗粒(NLP)并形成适合相干X射线衍射成像(CXDI)的微晶体的方法；John Spudich博士，他来自德克萨斯大学医学院。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School和Adam Cohen博士，他在哈佛大学的实验室最近证明了视紫红质可以用来监测活神经的跨膜电压。

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.