Genetic Analysis and Manipulation Core (GAEC)

摘要 遗传分析和编辑核心（GAEC）提供具有成本效益的，最先进的分子工具， 为IDDRC研究者提供咨询/生物信息学服务，1）。在所有尺度上测序DNA，2）。发展 用于人类神经发育疾病的细胞和动物模型的基因编辑策略，以及3）。 正常和病变细胞和组织中基因表达的定量分析。GAEC提供服务 通过其遗传分析组（GAG）和基因编辑组（GEG）。总检察长提供桑格 测序以验证质粒构建体、测序PCR产物以及所有需要 DNA测序，以及一整套用于基因组水平分析的检测和工具，从高通量开始 DNA纯化、定量和质量控制分析，分析性DNA检测，包括低， 中等和高通量形式，桑格和下一代测序（NGS），结构变体 检测和低、中和高通量基因表达研究，包括单细胞转录组学。 还提供生物信息学支持。GEG可开发人类IDDs的动物模型 利用最新的基因靶向方法，专门从事小鼠基因组的基因编辑， CRISPR/Cas9（规则间隔短回文重复序列）/（CRISPR相关蛋白），其 使得能够在单个基因组中产生具有点突变、插入或缺失的新型基因编辑小鼠系。 步还提供冷冻保存服务以及关于编辑策略和基因分型的建议。 将这些重叠的分析和编辑技术整合到一个核心设施中， 多个调查员允许单个调查员获得专门技术，并使用这些技术， 鉴定IDDRC相关突变，在小鼠和细胞系中模拟这些突变，并测量基因的变化， 在组织中或在单细胞水平表达。将两个已有的内核组合成一个集成的 核心使其重叠的活动更好地协调一致，并提供更全面和完整的 为IDDRC调查人员提供服务。

ABSTRACT The Genetic Analysis and Editing Core (GAEC) provides cost-effective, state-of-the-art molecular tools and consulting/bioinformatic services to IDDRC investigators for 1). Sequencing DNA at all scales, 2). Development of gene editing strategies for cellular and animal models of human neurodevelopmental disease, and 3). Quantitative analyses of gene expression in normal and diseased cells and tissues. GAEC offers services through its Genetic Analysis Group (GAG) and the Gene Editing Group (GEG). The GAG offers Sanger sequencing to validate plasmid constructs, sequence PCR products and for all molecular experiments requiring DNA sequencing, and a full suite of assays and tools for genomic-level analysis, starting with high throughput DNA purification, quantification and quality control assays, analytical DNA testing including genotyping in low, medium and high throughput formats, Sanger and next generation sequencing (NGS), structural variant detection, and low, medium and high throughput gene expression studies including single cell transcriptomics. Bioinformatic support is also provided. The GEG enables the development of animal models of human IDDs utilizing the latest gene targeting approaches and specializes in gene editing of the mouse genome via CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeat)/(CRISPR associated protein) which enables generation of novel gene-edited mouse lines with point mutations, insertions or deletions in a single step. A cryopreservation service is also offered as well as advice on editing strategies and genotyping. Consolidation of these overlapping analytic and editing technologies within a single core facility serving multiple investigators allows individual investigators access to specialized techniques and to use these to identify IDDRC related mutations, model these in mice and cell lines, and measure changes in gene expression in tissues or at a single cell level. The combination of two pre-existing cores into a single integrated core brings their overlapping activities into better alignment and offers a more comprehensive and complete service to IDDRC investigators.