Generation and initial charcterization of a mouse with floxed miR-155 for conditi

具有 floxed miR-155 条件的小鼠的生成和初步表征

• 批准号: 7990884
• 负责人: THOMAS MOELLER
• 金额: $ 7.8万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7990884
• 关键词: 3' Untranslated Regions; Actins; Area; Astrocytes; Cells; Development; Excision; Gene Expression Regulation; Generations; Genetic; Genetic Recombination; Goals; Infarction; Injury; Knock-out; Knockout Mice; Mediating; Memory; Messenger RNA; MicroRNAs; Microglia; Middle Cerebral Artery Occlusion; Modeling; Molecular; Multiple Sclerosis; Mus; Neurons; Nucleotides; Pathway interactions; Proteins; RNA; Regulation; Reperfusion Therapy; Role; Services; Signal Transduction; Site; Stroke; Synapses; Therapeutic Intervention; Translations; Trauma; Untranslated Regions; cell type; central nervous system injury; immune function; public health relevance; research study

DESCRIPTION (provided by applicant): Recent progress in genetics has identified a new class of RNA molecules called microRNAs (miRNA) involved in gene regulation. miRNAs are single-stranded RNA molecules of ~22 nucleotides complementary to a site in the 3' untranslated region (UTR) of mRNAs. The annealing of the miRNA to the mRNA inhibits protein translation and sometimes facilitates cleavage of the mRNA. This regulation adds an unexpected layer of complexity to the classic "linear" concept of DNA->mRNA->protein. Fast paced progress in the recent years showed the importance of miRNA in regulating neuronal and immune functions. In the CNS, miRNAs have been implicated in synaptic development and memory formation. We have found that constitutive knock out of miR-155 (miR-155-/-) increases the infract area in the mouse middle cerebral artery occlusion/reperfusion (MCAO/R) model by ~50%. However, miR-155 is expressed in the CNS in by at least astrocytes and microglia. The constitutive knock out of miR-155-/- makes it impossible to determine which cell type is responsible for the substantial increase in infarct area. We therefore wish to generate a conditional knock out model using the Cre/loxP approach. Using this well established approach we will be able to cross the floxed miR-155 (miR-155fl/fl) with readily available Cre-driver mice for astrocytes and microglia to generate cell specific knock out mice. The long-term goal of this project is to generate conditional knock outs of miR-155 to investigate the role of cell-specific miR-155 in CNS injury such as ischemic injury. We believe that the molecular mechanisms identified in subsequent experiments using this model may indentify new pathways for therapeutic intervention in CNS injuries such as stroke. PUBLIC HEALTH RELEVANCE: The broad, long-term goal of this project is to understand the role of miR-155 in the activation of astrocytes microglial cells. We believe that the signal transduction mechanisms identified through our experiments may constitute new targets for therapeutic intervention in CNS injuries associated with glial activation such as trauma, multiple sclerosis or stroke.

描述(申请人提供)：遗传学的最新进展发现了一类新的RNA分子，称为microRNAs(MiRNA)，参与基因调控。MiRNAs是一种由~22个核苷酸组成的单链RNA分子，与mRNAs的3‘非翻译区(UTR)上的一个位点互补。MiRNA对mRNA的热处理抑制了蛋白质的翻译，有时还促进了mRNA的切割。这一规定为DNA--mRNA-蛋白质这一经典的“线性”概念增添了一层意想不到的复杂性。近年来的快速发展表明了miRNA在调节神经元和免疫功能方面的重要性。在中枢神经系统中，miRNAs与突触发育和记忆形成有关。我们发现，在小鼠大脑中动脉阻塞/再灌注(MCAO/R)模型中，miR-155(miR-155-/-)的结构性敲除使梗塞面积增加约50%。然而，miR-155在中枢神经系统中至少由星形胶质细胞和小胶质细胞表达。MiR-155-/-的结构性敲除使得无法确定哪种细胞类型是导致梗死面积大幅增加的原因。因此，我们希望使用CRE/loxP方法生成一个条件敲除模型。使用这一成熟的方法，我们将能够将已分离的miR-155(miR-155fl/fl)与现成的Cre驱动小鼠杂交，以获得星形胶质细胞和小胶质细胞，从而产生细胞特异性基因敲除小鼠。该项目的长期目标是产生有条件的miR-155基因敲除，以研究细胞特异性miR-155在中枢神经系统损伤(如缺血性损伤)中的作用。我们相信，在随后使用该模型的实验中确定的分子机制可能为中风等中枢神经系统损伤的治疗干预确定新的途径。 公共卫生相关性：该项目的广泛、长期目标是了解miR-155在星形胶质细胞小胶质细胞激活中的作用。我们认为，通过我们的实验确定的信号转导机制可能构成与神经胶质细胞激活相关的中枢神经系统损伤治疗干预的新靶点，如创伤、多发性硬化症或中风。