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Optical sensing of voltage, pH, and small molecules using microbial rhodopsins

使用微生物视紫红质对电压、pH 和小分子进行光学传感

基本信息

批准号：
7981713
负责人：
Adam Ezra Cohen
金额：
$ 252万
依托单位：
HARVARD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-30 至 2015-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (Provided by the applicant) Abstract: Our goal is to introduce a new class of genetically encoded optical indicators, based on the huge diversity of environmentally sensitive spectral shifts that occur naturally in microbial rhodopsin proteins. We recently created a microbial rhodopsin-based fluorescent indicator of pH, with a sensitive range from pH 6.8 to 8.8. We have preliminary results on a microbial rhodopsin-based indicator of membrane potential, which shows greater sensitivity than any existing optical sensor of membrane potential. Just as GFP revolutionized biology through its ability to track the positions of proteins in cells, we believe that microbial rhodopsins will have a broad impact through their ability to transduce the physical and chemical environment into an optical signal. Sensing voltage is our first target. Neuroscientists have long dreamed of a genetically encoded sensor that gives an optical signal in response to a change in membrane potential, with the goal of imaging electrical activity of neurons in vivo. Such a molecule could also be used to probe membrane potentials in mitochondria, cardiac cells, or in other non-neuronal cells. Our strategy is completely different from previous approaches to optical voltage sensing, and has already shown promising results. The technical implementation involves a) protein engineering and directed evolution to optimize an electrochromic response, and b) design and construction of an ultrasensitive laser imaging system capable of detecting this response in living cells. The methodology developed for sensing pH and voltage will later be applied to other sensing modalities, such as chloride and membrane tension. Public Health Relevance: We are working to develop a new class of molecules that allow us to see changes in voltage or pH inside of single cells. Neurons use voltage to communicate, so the ability to see neuronal activity will provide insights into brain function.

描述（由申请人提供）

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（4）

Screening fluorescent voltage indicators with spontaneously spiking HEK cells.

筛选荧光电压指示器自发尖峰HEK细胞。

DOI：
10.1371/journal.pone.0085221
发表时间：
2013
期刊：
PloS one
影响因子：
3.7
作者：
Park J;Werley CA;Venkatachalam V;Kralj JM;Dib-Hajj SD;Waxman SG;Cohen AE
通讯作者：
Cohen AE

Photostick: a method for selective isolation of target cells from culture.

DOI：
10.1039/c4sc03676j
发表时间：
2015-03
期刊：
Chemical science
影响因子：
8.4
作者：
Chien MP;Werley CA;Farhi SL;Cohen AE
通讯作者：
Cohen AE

Convex lens-induced confinement for imaging single molecules.

DOI：
10.1021/ac101041s
发表时间：
2010-07-15
期刊：
ANALYTICAL CHEMISTRY
影响因子：
7.4
作者：
Leslie, Sabrina R.;Fields, Alexander P.;Cohen, Adam E.
通讯作者：
Cohen, Adam E.

Optogenetic Approaches to Drug Discovery in Neuroscience and Beyond.

DOI：
10.1016/j.tibtech.2017.04.002
发表时间：
2017-07
期刊：
Trends in biotechnology
影响因子：
17.3
作者：
Zhang H;Cohen AE
通讯作者：
Cohen AE

All-Optical Electrophysiology for High-Throughput Functional Characterization of a Human iPSC-Derived Motor Neuron Model of ALS.

DOI：
10.1016/j.stemcr.2018.04.020
发表时间：
2018-06-05
期刊：
Stem cell reports
影响因子：
5.9
作者：
Kiskinis E;Kralj JM;Zou P;Weinstein EN;Zhang H;Tsioras K;Wiskow O;Ortega JA;Eggan K;Cohen AE
通讯作者：
Cohen AE