GENERATION OF AN INDUCIBLE SYSTEM IN THE UTERINE STROMA FOR IMPLANTATION STUDIES

用于植入研究的子宫间质中诱导系统的生成

• 批准号: 8358586
• 负责人: Liang Ma
• 金额: $ 23.64万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-03 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358586
• 关键词: Accounting; Alleles; Binding; Chimera organism; Complex; Decidual Cell Reactions; Development; Doxycycline; Embryo; Embryo Loss; Embryonic Development; Event; Failure; Fertilization in Vitro; Gene Deletion; Gene Expression; Gene Targeting; Generations; Genes; Genetic; Genetic Recombination; Growth Factor; Hormonal; Human; Implant; Infertility; Knock-in Mouse; Knock-out; Knowledge; Lead; Mediating; Molecular; Mouse Strains; Mus; Nature; Organogenesis; Partner in relationship; Pathway interactions; Pharmaceutical Preparations; Phase; Placentation; Pregnancy; Procedures; Process; Production; Reporter; Research; Research Personnel; Southern Blotting; System; Technology; Tetracyclines; Tissues; Uterus; Validation; blastocyst; carcinogenesis; design; embryonic stem cell; failure Implantation; field study; homologous recombination; implantation; improved; intercellular communication; interest; natural Blastocyst Implantation; overexpression; postnatal; reproductive; tool; uterine receptivity; Accounting; Alleles; Binding; Chimera organism; Complex; Decidual Cell Reactions; Development; Doxycycline; Embryo; Embryo Loss; Embryonic Development; Event; Failure; Fertilization in Vitro; Gene Deletion; Gene Expression; Gene Targeting; Generations; Genes; Genetic; Genetic Recombination; Growth Factor; Hormonal; Human; Implant; Infertility; Knock-in Mouse; Knock-out; Knowledge; Lead; Mediating; Molecular; Mouse Strains; Mus; Nature; Organogenesis; Partner in relationship; Pathway interactions; Pharmaceutical Preparations; Phase; Placentation; Pregnancy; Procedures; Process; Production; Reporter; Research; Research Personnel; Southern Blotting; System; Technology; Tetracyclines; Tissues; Uterus; Validation; blastocyst; carcinogenesis; design; embryonic stem cell; failure Implantation; field study; homologous recombination; implantation; improved; intercellular communication; interest; natural Blastocyst Implantation; overexpression; postnatal; reproductive; tool; uterine receptivity

DESCRIPTION (provided by applicant): Human infertility is a global problem and failure of embryo implantation accounts for a significant percentage of pregnancy failure during both natural pregnancy and in vitro fertilization procedures. Implantation is an extremely complicated process requiring precisely controlled hormonal, growth factor signaling and cell-cell contacts which coordinate interactions between the competent blastocysts and the receptive uterus. In the past two decades we have greatly improved our knowledge on this subject by using sophisticated mouse genetics. However, our understanding of implantation is still rudimentary. Implantation research including the study of uterine receptivity and decidualization has benefited greatly from the Cre/LoxP technology which allows functional study of many genes in a tissue specific manner. Although inducible Cre/LoxP systems have been widely used in other fields of studies, their use in implantation studies has been limited by the steroidal nature of most of the inducers which interferes with the implantation process. The currently used Pgr-Cre and Amhr2-Cre lines are not inducible and each has their own limitations for implantation studies. Thus there is urgent need to develop an inducible tissue-specific Cre system for conditional deletion of genes in the uterus during implantation. In this proposal, we propose to knock rtTA into the endogenous Gli2 locus to generate a tetracycline-inducible line which in corporation with tetO-Cre can drive inducible Cre expression in the uterine stroma during implantation. In Aim I we will use BAC recombineering to generate a knock-in construct which will be used for gene targeting in ES cells and eventually germline chimera production. In Aim II, we will assess the usefulness of this knock-in allele in implantation studies. This mouse strain should be a valuable tool for researchers studying implantation as well as embryonic or postnatal organogenesis and carcinogenesis. PUBLIC HEALTH RELEVANCE: This project proposes to generate a tissue-specific inducible Cre system in the uterine stroma which can be used to knockout or overexpress gene of interest in the peri-implantation uterine stroma. This strain of mice will be an invaluable tool for reproductive biologists studying uterine receptivity, decidualization and placentation.

描述（由申请人提供）：人类不育症是一个全球问题，胚胎植入的失败占自然怀孕和体外受精程序期间妊娠失败的很大一部分。植入是一个极其复杂的过程，需要精确控制的激素，生长因子信号传导和细胞 - 细胞接触，该过程协调了既定的胚泡和接受性子宫之间的相互作用。在过去的二十年中，我们通过使用复杂的小鼠遗传学大大提高了我们对该主题的知识。但是，我们对植入的理解仍然是基本的。植入研究（包括对子宫接受性和脱义化的研究）从CRE/LOXP技术中受益匪浅，该技术允许以组织特定方式对许多基因进行功能研究。尽管可诱导的CRE/LOXP系统已被广泛用于其他研究领域，但它们在植入研究中的使用受到了大多数诱导剂的类固醇性质的限制，这些诱导剂会干扰植入过程。当前使用的PGR-CRE和AMHR2-CRE线不可诱导，并且每种植入研究都有自己的局限性。因此，迫切需要开发诱导的组织特异性CRE系统，以在植入过程中有条件地缺失子宫中基因的条件缺失。在此提案中，我们建议将RTTA撞到内源性GLI2基因座中，以产生四环素诱导的线，在使用TETO-CRE的公司中，在植入过程中，该线路可以驱动子宫基质中的诱导型CRE表达。在目的中，我将使用BAC重新组合来产生敲入构建体，该结构将用于ES细胞中的基因靶向，并最终用于生殖线嵌合体的产生。在AIM II中，我们将评估该敲门等位基因在植入研究中的有用性。该小鼠菌株应该是研究植入以及胚胎或产后器官发生和致癌作用的有价值的工具。 公共卫生相关性：该项目提议在子宫基质中生成组织特异性诱导的CRE系统，该系统可用于敲除或过表达子宫周期基质中感兴趣的基因。这种小鼠的菌株将是研究子宫接受能力，c骨化和胎盘的生殖生物学家的宝贵工具。