Fluorescent protein biosensors applied to microphysiological systems.
Fluorescent protein biosensors applied to microphysiological systems.
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DOI：
10.1177/1535370215584934
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发表时间：
2015-06
期刊：
影响因子：
--
通讯作者：
Taylor DL
中科院分区：
文献类型：
--
作者：
Senutovitch N;Vernetti L;Boltz R;DeBiasio R;Gough A;Taylor DL
本文讨论了荧光作为体外研究活细胞和组织的工具的发展，以及荧光蛋白生物传感器在微生理系统(MPS)中的当前作用。FPB允许实时测量正常和扰动的细胞分析系统和微生理系统中涉及的目标细胞事件的时间和空间动力学。FPB由荧光模拟细胞化学(FAC)发展而来，FAC允许测量用环境不敏感的荧光染料共价标记的纯化蛋白质的动力学，然后结合到活细胞中，以及一大批可扩散的荧光探针，这些荧光探针是为测量活细胞中的环境变化而设计的。与此同时，发展了一系列的荧光显微镜方法来测量标记细胞的化学和分子活性，包括比率成像、荧光寿命、全内反射、3D成像(包括超分辨率)以及高含量筛选(HCS)。FPBS由FAC进化而来，通过将环境敏感的荧光染料与蛋白质结合起来，以监测特定的生理事件，如翻译后修饰、代谢物的产生、各种离子浓度的变化以及蛋白质与细胞内特定大分子在时间和空间上的动态相互作用。最初的FPB涉及到荧光染料的工程，当共价结合到目标蛋白质的特定结构域上时，可以感觉到特定的活性。随后发展的荧光蛋白(FP)，如绿色荧光蛋白(GFP)，极大地加快了研究活细胞的采用，因为蛋白质的遗传“标记”成为一种相对简单的方法，允许分析广泛的蛋白质的时空动态。研究人员随后针对环境敏感性设计了FP的荧光特性，当与目标蛋白质/肽结合时，创建了新一代FPB。给出了在MPS中有用的FPBs的实例，包括设计、测试和在肝脏MPS中的应用。
This mini-review discusses the evolution of fluorescence as a tool to study living cells and tissues in vitro and the present role of fluorescent protein biosensors (FPBs) in microphysiological systems (MPS). FPBs allow the measurement of temporal and spatial dynamics of targeted cellular events involved in normal and perturbed cellular assay systems and microphysiological systems in real-time. FPBs evolved from fluorescent analog cytochemistry (FAC) that permitted the measurement of the dynamics of purified proteins covalently labeled with environmentally insensitive fluorescent dyes and then incorporated into living cells, as well as a large list of diffusible fluorescent probes engineered to measure environmental changes in living cells. In parallel, a wide range of fluorescence microscopy methods were developed to measure the chemical and molecular activities of the labeled cells, including ratio imaging, fluorescence lifetime, total internal reflection, 3D imaging, including super-resolution, as well as high content screening (HCS). FPBs evolved from FAC by combining environmentally sensitive fluorescent dyes with proteins in order to monitor specific physiological events such as post-translational modifications, production of metabolites, changes in various ion concentrations and the dynamic interaction of proteins with defined macromolecules in time and space within cells. Original FPBs involved the engineering of fluorescent dyes to sense specific activities when covalently attached to particular domains of the targeted protein. The subsequent development of fluorescent proteins (FPs), such as the green fluorescent protein (GFP), dramatically accelerated the adoption of studying living cells, since the genetic “labeling” of proteins became a relatively simple method that permitted the analysis of temporal-spatial dynamics of a wide range of proteins. Investigators subsequently engineered the fluorescence properties of the FPs for environmental sensitivity that, when combined with targeted proteins/peptides, created a new generation of FPBs. Examples of FPBs that are useful in MPS are presented, including the design, testing and application in a liver MPS.
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影响因子：
7.5
作者：
Bhushan A;Senutovitch N;Bale SS;McCarty WJ;Hegde M;Jindal R;Golberg I;Berk Usta O;Yarmush ML;Vernetti L;Gough A;Bakan A;Shun TY;DeBiasio R;Lansing Taylor D
通讯作者：
Lansing Taylor D
DOI：
10.1177/1535370214531872
发表时间：
2014-09
期刊：
Experimental biology and medicine (Maywood, N.J.)
影响因子：
--
作者：
Bale SS;Vernetti L;Senutovitch N;Jindal R;Hegde M;Gough A;McCarty WJ;Bakan A;Bhushan A;Shun TY;Golberg I;DeBiasio R;Usta BO;Taylor DL;Yarmush ML
通讯作者：
Yarmush ML
影响因子：
7
作者：
Cho SW;Kim S;Kim Y;Kweon J;Kim HS;Bae S;Kim JS
通讯作者：
Kim JS
影响因子：
56.9
作者：
Cole, NB;Smith, CL;LippincottSchwartz, J
通讯作者：
LippincottSchwartz, J
影响因子：
17.3
作者：
Gaj, Thomas;Gersbach, Charles A.;Barbas, Carlos F., III
通讯作者：
Barbas, Carlos F., III