Production of a humanized Ah receptor mouse line

人源化Ah受体小鼠系的生产

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要/摘要芳基烃受体（AHR）的生理功能是一个敏锐的领域，尤其是在其在肠道稳态，微生物组组成和免疫调节中的作用的术语。另外，没有胃肠道中的AHR表达导致对不利挑战的敏感性增加，例如柠檬酸啮齿动物暴露。也许这部分是由于AHR在IL-22中扮演的核心作用肠内的表达。我们发表了许多鉴定内源或微生物组的文章产生的AHR配体，其中许多人被发现是显着有效的激活剂与鼠标AHR相比，人类AHR。这些研究导致了人类的中心假设与小鼠AHR相比，AHR在肠道稳态中起着更大的作用。为了测试这个假设，我们需要开发合适的人源化AHR小鼠。因此，具体的目的是产生表达人AH受体的敲入小鼠。实际假设将在随后的赠款申请。小鼠线将由Cyagen生成；一家广泛的公司有产生敲门鼠的经验。我们方法的关键方面是使用密码子优化，并且次级RNA结构破坏了合成人AHR cDNA，该c cDNA表现出高度高4倍表达比野生型人AHR cDNA。敲入人类鼠标线将被描述为确定它相对于C57BL/6J小鼠表现出类似的AHR表达水平和模式。在此外，AHR靶基因的诱导水平适用于人AHR的配体特异性先前已建立。该人源化的AHR小鼠将对有关该作用的广泛研究有用 AHR在疾病过程中，并评估人类AHR作为治疗靶点。