A mouse model for genetic tracing to study stress responses

用于研究应激反应的基因追踪小鼠模型

基本信息

批准号：
8385998
负责人：
TSONWIN HAI
金额：
$ 22.88万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8385998
关键词：
A Mouse Acute Address Affect Alleles Area Biochemical Biological Cell Separation Cells Chronic Chronic stress Color Core Facility DNA Development Diet Disease Environment Erythrocytes Event Exploratory/Developmental Grant Fatty acid glycerol esters Fluorescence Future Gene Expression Genes Genetic Goals Green Fluorescent Proteins Half-Life Harvest Health Heart Human Injection of therapeutic agent Injury Label Lipopolysaccharides Liver Lung Malignant Neoplasms Modeling Molecular Mus Myocardial Infarction National Institute of Environmental Health Sciences Nerve Degeneration Nuclear Obesity Organ Play Process Production Proteins Reagent Reporter Research Research Personnel Risk Role Signal Transduction Site Sorting - Cell Movement Stress System Tamoxifen Testing Time Tissue Harvesting Toxin Transgenes Transgenic Mice Transgenic Organisms acute stress biological adaptation to stress cellular imaging design gene environment interaction innovation interest mouse model novel recombinase response tool web site

项目摘要

DESCRIPTION (provided by applicant): The environment to which humans are exposed often contains toxins and stress factors, which induce a variety of biological responses. These are gene-directed active processes; thus, how the cells respond and how genes and environments interact play an important role in disease development. The goal of this R21 is to establish a mouse model to study the biological responses to various environmental stresses. Designs and Utilities: The design is such that it will allow the researchers to trace and study both the cells under stress at the time of harvesting the tissues and the cells that were under stress in their past - long before harvesting. The mice will carry two transgenic alleles. 1. Stress-inducible Cre*: encodes a recombinase Cre fusion (Cre* for short), whose expression is turned on by a broad spectrum of stress signals, and 2. ROSA-Reporter: encodes Yellow Fluorescent Protein (YFP) preceded by Lox-STOP-Lox (LSL). The production of YFP is blocked by LSL and is possible only if the STOP is removed by active Cre*. This event physically and thus permanently alters the DNA, resulting in YFP production (green) to forever "mark" the cells. This allows one to "trace" the cells and determine their "fate. The Cre* is fluorescent (red); thus, the cells can be labeled red, if they are actively under stres (producing Cre*). With proper designs, one can distinguish cells that were once under stress in their past from those currently under stress. The fluorescent colors allow not only cell imaging but also cell isolation (by sorting) for biochemical and molecular analyses. Aim 1: To generate and characterize the stress-inducible Cre* transgenic mice. The transgenic construct will be generated using BAC recombineering and injected into pronucleus to make the mice. The mice will then be characterized for the expression and half-life of Cre* under acute and chronic stress paradigms. Aim 2: To generate and characterize the double transgenic mice. The stress-inducible Cre* mice will be crossed with the YFP reporter mice (available) to generate the double transgenic mice. The mice will then be analyzed to determine how tight the system is and how efficient the active Cre* is in removing the STOP. Innovation and Significance: The mouse model is novel for its ability to trace the fate of cells under various stresses, such as those induce liver injury, heart attack, neuronal degeneration, and cancer development. In addition, the proposed mouse model allows researchers to address issues regarding the transition from acute to chronic (or repeated) injuries¿an important but not well understood issue. The broad inducibility of Cre* makes the model an excellent tool to study environmentally-induced diseases - a special emphasis of NIEHS. PUBLIC HEALTH RELEVANCE: The goal of this R21 is to establish a novel mouse model to obtain new understanding of environmentally- induced diseases. The design is such that it will allow the researchers to trace and study both the cells under stress at the time of harvesting the tissues and the cells that were under stress in their past-long before harvesting. The successful establishment of the model will allow us to trace the fate of cells under a variety of stresses, such as those induce liver injury, heart attack, neuronal degeneration, and cancer development.

描述（由申请人提供）：暴露于人类的环境通常含有毒素和压力因素，从而诱导各种生物学反应。这些是基因指导的主动过程；因此，细胞的反应以及基因和环境如何相互作用在疾病发展中起重要作用。该R21的目的是建立小鼠模型来研究对各种环境应力的生物学反应。设计和实用程序：设计将使研究人员能够在收获时追踪和研究两个细胞。在收获之前很长时间之前，组织和细胞承受压力。小鼠将携带两个转基因等位基因。 1。应力诱导的CRE*：编码重组酶CRE融合（简称CRE*），其表达是通过广泛的应力信号打开的，2。Rosa-Reporter：编码黄色荧光蛋白（YFP），由Lox-Stop-lox（LSL）（LSL）。 YFP的产生被LSL阻止，并且只有在通过Active CRE*去除停止时才有可能。该事件在物理上并因此永久改变了DNA，从而导致YFP产生（绿色）永远“标记”细胞。这使人们可以“追踪”细胞并确定其“命运。cre*是荧光（红色）；因此，如果它们在压力下积极压力（产生CRE*），则可以将细胞标记为红色。通过适当的设计，可以区分曾经在压力下的压力的细胞与当前压力下的压力区分开，而静脉颜色不仅允许单元格的目的。生成和诱导的转基因小鼠将使用BAC重组产生转基因构建体，并将其注射到pronuce中，以使小鼠的表达和半生命的表达方式。然后，将分析小鼠（可用的）。从急性到慢性（或重复）伤害的过渡是一个重要但尚不清楚的问题。公共卫生相关性： R21的目的是建立一种新型的小鼠模型，以获得对环境诱发疾病的新理解。该设计使研究人员可以在收获组织和在收获前长时间承受压力的细胞时追踪和研究压力下的细胞。该模型的成功建立将使我们能够在各种压力下追踪细胞的命运，例如影响肝损伤，心脏病发作，神经元发育和癌症发育。