Regulation of singular expression in Trace Amine-Associated Receptors

痕量胺相关受体中奇异表达的调节

• 批准号: 9420717
• 负责人: Ami Shah
• 金额: $ 0.24万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-31 至 2020-01-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9420717
• 关键词: Alleles; Amination; Amines; Binding Sites; CRISPR/Cas technology; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Distal; Elements; Enhancers; Environment; Epigenetic Process; Family; Gene Cluster; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Gene Targeting; Gene Transfer Techniques; Genes; Genetic; Genome; Genomics; Goals; Heterochromatin; Human; Hydrogen; Individual; Light; Mammals; Mediating; Modeling; Molecular; Mus; Nasal Epithelium; Nasal cavity; Nervous system structure; Neurons; Odorant Receptors; Odors; Olfactory Pathways; Phosphorus; Phylogenetic Analysis; Play; Population; Probability; Quality of life; Receptor Gene; Regulation; Regulatory Element; Reporting; Research Personnel; Role; Site; Smell Perception; Tellurium; Testing; Time; Training; Transcription Initiation Site; Transgenes; Work; cell type; cis acting element; epigenetic regulation; experimental study; gene function; genetic approach; genetic element; genome editing; histone modification; homeodomain; insight; interest; olfactory receptor; olfactory sensory neurons; prevent; promoter; receptor; receptor expression; transcriptome sequencing

Project Summary Olfaction is essential for detecting volatile molecules in the environment, is crucial for the survival of many vertebrate species, and contributes significantly to quality of life in humans. In mammals, odors are detected by neurons in the main olfactory system using a large family odorant receptor (OR) genes. These genes show an unusual form of gene regulation—each olfactory neuron expresses a single allele of a single OR gene. The mechanism underlying this random monoallelic (or “monogenic”) expression is not understood, but likely involves the interaction of proximal and distal cis-acting elements, as well as epigenetic mechanisms. Mammals also retain a smaller family of main olfactory receptors, the Trace Amine Associated Receptors (TAARs). The TAARs are also subject to monogenic expression. However, recent data have uncovered both similarities and differences between OR and TAAR gene expression—some of which raise fundamental questions about current models of OR choice. Reconciling these differences and shedding light on TAAR expression are both critical to our understanding of gene choice mechanisms in olfaction. The goal of this proposal is to characterize for the first time genetic elements that are involved in monogenic expression of TAAR genes. I have identified two well-conserved, putative regulatory elements in the TAAR gene cluster. I propose to use advanced mouse genetic approaches to test whether and how these elements contribute to TAAR gene choice. Specific Aim 1 will use transgenesis and gene targeting via CRISPR/Cas9 to test whether one or both of these elements is sufficient to drive singular expression in specific populations of olfactory sensory neurons that normally express TAARs. Specific Aim 2 will use genome editing via CRISPR/Cas9 to determine if one or both of these elements is necessary for singular expression of TAAR genes. These experiments provide necessary first steps towards characterizing the similarities and differences between OR and TAAR gene choice mechanisms. Such comparisons will provide insight into the basic mechanisms of monoallelic gene expression and cell type diversity in the mammalian nervous system.

项目摘要 嗅觉对于检测环境中挥发性分子至关重要，对于许多人的存活至关重要 脊椎动物物种，并为人类的生活质量做出了重大贡献。在哺乳动物中，检测到气味 由主嗅觉系统中的神经元使用大型家族气质受体（OR）基因。这些基因显示 基因调节的一种不寻常的形式 - 嗅觉神经元表达单个或基因的单个等位基因。这 不了解这种随机的单相（或“单基因”）表达的机制，但可能 涉及近端和远端顺式作用元件以及表观遗传机制的相互作用。 哺乳动物还保留了较小的主嗅觉受体家族，痕量胺相关受体 （taars）。 TAAR还受到单基因表达的影响。但是，最近的数据发现了这两者 OR和TAAR基因表达之间的相似性和差异 - 其中一些提高了基本 有关当前或选择模型的问题。核对这些差异，并在taar上散发灯光 表达对于我们对嗅觉中基因选择机制的理解至关重要。 该提议的目的是在首次参与的遗传因素表征 TAAR基因的单基因表达。我已经确定了两个保守的，推定的监管元素 TAAR基因簇。我建议使用先进的鼠标遗传方法来测试这些方法是否以及如何 元素有助于TAAR基因选择。特定目标1将使用转基因和基因靶向通过 CRISPR/CAS9测试其中一个或两个元素是否足以在特定的 通常表达TAAR的嗅觉感觉神经元的种群。特定目标2将使用基因组编辑 通过crispr/cas9来确定这些元素中的一个或两个是taar的单数表达 基因。这些实验提供了表征相似性和差异的必要第一步 在OR和TAAR基因选择机制之间。这种比较将提供对基本的见解 哺乳动物神经系统中单相基因表达和细胞类型多样性的机制。