Generation and Validation of a Conditional Circadian Reporter Mouse

条件昼夜节律报告小鼠的生成和验证

基本信息

批准号：
9379636
负责人：
DAVID Raymond WEAVER
金额：
$ 20.94万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9379636
关键词：
Address Albumins Alleles Alpha Cell Animals Argipressin Automobile Driving Behavior Behavioral Binding Proteins Bioluminescence Brain Brain imaging Brain region Cardiovascular Diseases Cell Line Cells Characteristics Chimeric Proteins Circadian Rhythms Communities Complement Complex Darkness Dependence Dissection Dopamine D1 Receptor Elements Ensure Enterobacteria phage P1 Cre recombinase Environment Event Feedback Funding Gene Expression Generations Genes Genetic Genetic Transcription Goals Health Hepatocyte Hour Image In Vitro Incidence Individual Islets of Langerhans Kidney Knock-in Mouse Letters Light Liver LoxP-flanked allele Luciferases Malignant Neoplasms Masks Messenger RNA Methods Modification Molecular Monitor Mus Nature Neurons Obesity Organ Outcome Output Pancreas Periodicity Peripheral Phenotype Photons Physiological Physiology Play Population Proteins Rattus Reporter Reporter Genes Reporting Research Research Personnel Rest Retina Rodent Role Signal Transduction Slice Source Stroke Submandibular gland Tamoxifen Testing The Jackson Laboratory Time Tissues Transgenic Mice United States National Institutes of Health Validation awake base cell type circadian pacemaker design and construction in vivo in vivo bioluminescence imaging interest islet light emission luciferin mouse model novel promoter repository shift work suprachiasmatic nucleus virtual whole animal imaging

项目摘要

Most aspects of physiology have 24-hour rhythmicity. The master clock regulating circadian physiology and behavior is located in the brain, in the suprachiasmatic nuclei (SCN). Other tissues also generate rhythms of gene expression and have near-24-hr rhythms in function. An enduring question in the field is how the rhythmicity in different cells and tissues is coordinated. There is a significant unmet need for lines of mice that allow monitoring of circadian rhythms in specific tissues and cell types. The objective of this proposal is to generate and validate two novel lines of mice expressing luciferase in a rhythmic manner. In one line of mice, luciferase expression will be conditional on Cre recombinase and in the other line, luciferase is expressed in a non-conditional way. These lines will be generated by modifications of the highly rhythmic and widely expressed gene encoding albumin D-element binding protein (DBP). Our proposed studies will characterize these reporter lines for utility in generating bioluminescence rhythms in a variety of tissues, including liver, pancreas and brain, will verify the Cre-dependence of luciferase activity in the Cre-dependent line, and will verify that the genetic targeting event does not alter behavioral or molecular rhythmicity of the animals bearing it, as necessary for this to be a useful circadian reporter. We anticipate these lines of mice will be valuable to many investigators studying circadian rhythms in brain and peripheral tissues, and will facilitate studies of the desynchrony among tissues that occurs following shifts of the light-dark cycle. Once validated, these lines of mice will be distributed to qualified investigators directly and via the Jackson Laboratories repository.

生理学的大多数方面都有24小时的节律性。调节昼夜节律的主时钟生理和行为位于大脑的视交叉上核（SCN）。其他组织也会产生基因表达节律，并且具有近 24 小时的功能节律。一个该领域长期存在的问题是不同细胞和组织的节律性如何协调。对于能够监测昼夜节律的小鼠品系的需求存在显着的未满足的需求特定的组织和细胞类型。该提案的目标是生成并验证两个以有节奏的方式表达荧光素酶的新小鼠系。在一系列小鼠中，荧光素酶表达将以 Cre 重组酶为条件，在另一株系中，荧光素酶表达于一种无条件的方式。这些线条将通过修改高度节奏和广泛表达的编码白蛋白 D 元件结合蛋白 (DBP) 的基因。我们提出的研究将描述这些报告线在产生生物发光节律方面的效用多种组织，包括肝脏、胰腺和大脑，将验证荧光素酶的 Cre 依赖性 Cre 依赖性系中的活性，并将验证遗传靶向事件不会改变携带它的动物的行为或分子节律，这是有用的必要条件昼夜节律记者。我们预计这些小鼠品系对许多研究人员来说都是有价值的研究大脑和周围组织的昼夜节律，并将促进对明暗周期变化后发生的组织间不同步。一旦验证通过，这些小鼠品系将通过 Jackson 直接分发给合格的研究人员实验室存储库。