Production of a humanized Ah receptor mouse line

人源化Ah受体小鼠系的生产

• 批准号: 9207268
• 负责人: Gary H. Perdew
• 金额: $ 7.86万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9207268
• 关键词: AHR gene; Address; Applications Grants; Area; Aromatic Polycyclic Hydrocarbons; Aryl Hydrocarbon Receptor; Bacterial Artificial Chromosomes; Bicyclo Compounds; Chemicals; Citrobacter; Citrobacter rodentium; Codon Nucleotides; Communities; Complementary DNA; Data; Development; Diet; Dioxins; Disease; Drug Industry; Drug Targeting; Enzymes; Exhibits; Exons; Exposure to; Future; Gastrointestinal tract structure; Gene Targeting; Goals; Half-Life; Health; Homeostasis; Human; In Vitro; Indican; Indole-3-Carbinol; Indoles; Intestines; Investigation; Japan; Knock-in; Knock-in Mouse; Kynurenic Acid; Laboratories; Lead; Letters; Ligands; Liver; Lymphocyte; Lymphoid Follicle; Mediating; Metabolism; Mouse Strains; Mus; Pathway interactions; Pattern; Physiological; Play; Predisposition; Process; Production; Publishing; RNA; Receptor Activation; Reporting; Resistance; Role; Sodium Dextran Sulfate; Source; Specificity; Structure; System; Testing; Tetrachlorodibenzodioxin; Tissues; Toxic effect; Transgenic Mice; Translations; Western Blotting; Xenobiotics; aryl hydrocarbon receptor ligand; bacterial metabolism; experience; experimental study; gastrointestinal; gut microbiome; immune function; immunoregulation; interest; intraepithelial; microbiome; mouse model; overexpression; receptor expression; therapeutic target; transcription factor; γδ T cells

Project Summary/Abstract The physiological function of the aryl hydrocarbon receptor (AHR) is an area of keen interest, especially in terms of its role in gut homeostasis, microbiome composition, and immune regulation. In addition, absence of AHR expression in the gastrointestinal tract leads to increased susceptibility to an adverse challenge, such as citrobacter rodentium exposure. Perhaps this is due in part to the central role that the AHR plays in Il-22 expression within the gut. We have published a number of articles identifying endogenous or microbiome generated AHR ligands, many of which have been discovered to be dramatically more potent activators of the human AHR compared to mouse AHR. These studies have lead to the central hypothesis that the human AHR plays an increased role in gut homeostasis compared to the mouse AHR. In order to test this hypothesis, we need to develop a suitable humanized AHR mouse. Thus, the specific aim is to generate a knock-in mouse that expresses the human Ah receptor. The actual hypothesis would be addressed in a subsequent grant application. The mouse line will be generated by Cyagen; a company with extensive experience with generating knock-in mice. The key aspect of our approach is the use of a codon optimized and secondary RNA structure disrupted synthetic human AHR cDNA, which exhibits a 4-fold higher level of expression than wild-type human AHR cDNA. The knock-in human AHR mouse line will be characterized to establish that it exhibits a similar level and pattern of AHR expression relative to a C57BL/6J mouse. In addition, the level of induction of AHR target genes is appropriate for the ligand specificity of the human AHR previously established. This humanized AHR mouse will be useful for a wide range of studies of the role of the AHR in disease processes, as well as assessing the human AHR as a therapeutic target.

项目总结/摘要 芳香烃受体（AHR）的生理功能是一个非常感兴趣的领域，特别是在 就其在肠道稳态、微生物组组成和免疫调节中的作用而言。此外，缺乏 胃肠道中的AHR表达导致对不利挑战的易感性增加，例如 啮齿类柠檬酸杆菌暴露。也许这部分是由于AHR在伊尔-22中扮演的核心角色 肠道内的表达。我们已经发表了一些文章，确定内源性或微生物组 产生的AHR配体，其中许多已被发现是显着更有效的激活剂， 人AHR与小鼠AHR的比较。这些研究导致了一个中心假设，即人类 与小鼠AHR相比，AHR在肠道稳态中发挥更大的作用。为了测试这种 假设，我们需要开发合适的人源化AHR小鼠。因此，具体目标是生成一个 表达人Ah受体的基因敲入小鼠。实际的假设将在 随后的补助金申请。小鼠品系将由Cyagen生产，该公司拥有广泛的 基因敲入小鼠的经验。我们的方法的关键方面是使用密码子优化， 二级RNA结构被破坏的合成人AHR cDNA，其表现出4倍高的水平， 与野生型人AHR cDNA相比，将对敲入的人AHR小鼠系进行表征， 确定其表现出与C57 BL/6 J小鼠相似的AHR表达水平和模式。在 此外，AHR靶基因的诱导水平适合于人AHR的配体特异性。 以前建立的。这种人源化的AHR小鼠将可用于广泛的研究AHR的作用。 疾病过程中的AHR，以及评估人类AHR作为治疗靶点。