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.

项目概要 合成调控基因组学中心（SyRGe）的任务是开发和应用 对大基因位点进行协调改变的技术，广泛地促进功能研究 全基因组关联研究（GWAS）揭示的调控序列“成功”。 显着取代现有的调控基因组功能操作和评估技术 通过其对所谓“大 DNA”的组装和交付的关注，该补充请求提议： 利用我们在大 DNA 组装和交付方面最新的技术开发来解决大基因座的问题 CACNA1C，包含与各种神经精神疾病（包括精神分裂症）相关的热门药物， 双相情感障碍、重度抑郁症、ADHD 和 ASD 编码 L 型成孔亚基。 电压门控钙通道广泛表达于神经元、内分泌细胞、心肌细胞和 通过该通道的钙流入对于突触可塑性和基因表达至关重要。 调节、胰岛素分泌和肌肉收缩在这个补充请求中，我们建议组装和。 提供约 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 构建并传递包含风险或保护性等位基因的人类 CACNA1C 内含子 3，并区分 我们还将构建一系列变体以进行进一步的研究。 我们将评估分化神经元中的 Cacna1c mRNA 和 L 型钙电流。 本补充请求中提出的工作将有助于深入了解潜在的分子机制 多种神经精神疾病，也将为进一步产生人性化药物奠定基础 用于行为研究的 Cacna1c 小鼠模型。