Supplement for Center for Synthetic Regulatory Genomics: Building CACNA1C alleles associated with Neuropsychiatric Disorders

合成调控基因组学中心的补充：构建与神经精神疾病相关的 CACNA1C 等位基因

• 批准号: 10405299
• 负责人: Jef D BOEKE
• 金额: $ 38.26万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-12 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10405299
• 关键词: Affect; Alleles; Amino Acids; Attention deficit hyperactivity disorder; Behavior; Bipolar Disorder; Brain region; Calcium; Calcium Channel; Cardiac Myocytes; Cell Line; Cell Nucleus; Cell physiology; Cells; DNA; Data; Development; Disease; Disease susceptibility; Distal; Electrophysiology (science); Endocrine; Engineering; Enhancers; Exons; Foundations; Funding; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Polymorphism; Genome; Genomics; Haplotypes; Human; Human Genetics; Human Genome; Hybrids; Individual; Introns; Investigation; Length; Libraries; Linkage Disequilibrium; Major Depressive Disorder; Messenger RNA; Minisatellite Repeats; Molecular; Mouse Protein; Mus; Muscle Contraction; Neurons; Pan Genus; Phenotype; Physiology; Predisposition; Proteins; Quantitative Trait Loci; Regulatory Element; Risk; Schizophrenia; Series; Smooth Muscle; Synaptic plasticity; Tandem Repeat Sequences; Technology; Tissues; Untranslated RNA; Ursidae Family; Variant; Work; behavior test; behavioral study; brain cell; causal variant; cell type; experimental study; genetic variant; genome wide association study; genomic locus; humanized mouse; insight; insulin secretion; mouse model; neuropsychiatric disorder; neuropsychiatry; protective allele; protective effect; risk variant; synthetic construct; technology development; theories; tool; transcriptome sequencing; voltage

Project Summary The Center for Synthetic Regulatory Genomics (SyRGe) is tasked with development and application of technology for making coordinated changes to large gene loci, broadly enabling investigations into the function of regulatory sequences revealed by genome wide association study (GWAS) “hits”. The Center’s tools dramatically supersede present technologies for manipulation and assessment of regulatory genome function through its focus on assembly and delivery of so-called Big DNA. This supplement request proposes to leverage our most recent technology development in Big DNA assembly and delivery to tackle a large locus in CACNA1C, containing the top hits associated with various neuropsychiatric disorders, including schizophrenia, bipolar disorder, major depression, ADHD and ASD. CACNA1C encodes a pore-forming subunit of the L-type voltage-gated calcium channel and is widely expressed in neurons, endocrine cells, cardiomyocytes and smooth muscles. Calcium influx through the channel is critical for synaptic plasticity, gene expression regulation, insulin secretion and muscle contraction. In this supplement request, we propose to assemble and deliver the ~330 kb human CACNA1C intron 3 to replace the mouse endogenous Cacna1c intron 3. We will build and deliver human CACNA1C intron 3 that contains risk or protective alleles and differentiate the engineered mouse ESCs into neurons for initial phenotyping. We will also build a series of variants for further functional studies. We will assess the Cacna1c mRNA and L-type calcium current in differentiated neurons. The proposed work in this supplement request will enable insights into the molecular mechanisms underlying multiple neuropsychiatric disorders, and it will also serve as the foundation for further generation of humanized Cacna1c mouse models for behavioral studies.

项目摘要 合成调节基因组学中心（Syrge）的任务是开发和应用 用于对大基因局部进行协调更改的技术，从而广泛地研究了该功能 基因组广泛关联研究（GWAS）揭示的调节序列“ hits”。中心的工具 多龙取代了当前的操纵和评估调节基因组功能的技术 通过关注所谓的大型DNA的组装和交付。此补充请求提案 利用我们在大型DNA组装和交付方面的最新技术开发来解决一个大型轨迹 CACNA1C，包含与各种神经精神疾病有关的最高命中，包括精神分裂症， 躁郁症，重度抑郁症，ADHD和ASD。 cacna1c编码L型的孔形亚基 电压门控钙通道，并在神经元，内分泌细胞，心肌细胞和 平滑的肌肉。通过通道的钙影响对于突触可塑性，基因表达至关重要 调节，胰岛素分泌和肌肉收缩。在此补充要求中，我们建议组装和 传递〜330 kb的人Cacna1c内含子3以替换小鼠内源性Cacna1c内含子3。我们将 构建和交付包含风险或保护等位基因的人类CACNA1C内含子3 工程的小鼠移至神经元进行初始表型。我们还将构建一系列变体，以进一步 功能研究。我们将评估分化神经元中的CACNA1C mRNA和L型钙电流。 此补充要求中的拟议工作将使您能够洞悉分子机制 多种神经精神疾病，它也将成为进一步生成人性化的基础 CACNA1C小鼠模型用于行为研究。