Analysis and Bioengineering of Optogenetic Rhodopsins

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沿着利用先进的光谱和生物物理/生物工程技术。该研究的一个中心特征将是使用时间分辨FTIR差分光谱，超快激光光谱和共焦近红外拉曼技术来测量光遗传视紫红质分子的变化时，光激发的时间尺度为秒至飞秒。初步工作已经证明，各种光遗传视紫红质可以重组成蛋白脂质膜，包括nanolipidparticles和研究使用先进的光谱方法，以获得详细的分子信息，他们的功能。此外，光谱研究已扩展到活细胞，允许这些蛋白质在其天然环境中进行研究，以确定膜电位等因素的分子效应。该项目的合作者包括：麻省理工学院的艾德博伊登博士，被广泛认为是光遗传学的共同创始人之一，也是新神经光子技术的领先开发者; UCD的马特科尔曼博士，他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）的微晶的方法，美国国家科学院的约翰斯普迪奇博士，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，以及亚当·科恩博士，他在哈佛大学的实验室最近证明视紫红质可用于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.