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.

项目概要/摘要 芳烃受体（AHR）的生理功能是一个人们非常感兴趣的领域，特别是在 其在肠道稳态、微生物组成和免疫调节中的作用。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 啮齿动物柠檬酸杆菌暴露。也许这部分是由于 AHR 在 Il-22 中发挥的核心作用 肠道内的表达。我们发表了许多识别内源性或微生物组的文章 产生的 AHR 配体，其中许多已被发现是更有效的 AHR 激活剂 人类 AHR 与小鼠 AHR 的比较。这些研究得出了一个中心假设：人类 与小鼠 AHR 相比，AHR 在肠道稳态中发挥着更大的作用。为了测试这个 假设，我们需要开发一种合适的人源化AHR小鼠。因此，具体目标是生成一个 表达人类Ah受体的基因敲入小鼠。实际的假设将在 随后的拨款申请。鼠标线将由 Cyagen 生成；一家拥有广泛 产生基因敲入小鼠的经验。我们方法的关键方面是使用经过优化的密码子 二级RNA结构破坏了合成的人类AHR cDNA，其表现出4倍高的水平 表达量高于野生型人 AHR cDNA。敲入人 AHR 小鼠系的特征是 确定其表现出与 C57BL/6J 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类AHR的配体特异性 之前成立的。这种人源化 AHR 小鼠将有助于广泛研究 AHR 的作用 疾病过程中的 AHR，以及评估人类 AHR 作为治疗靶点。