GUDMAP2 - Production of Mouse Strains for Gene Anatomy of the Lower Urinary Tract

描述(申请人提供)：建议的研究计划是对RFA-DK-11-001，“生殖器解剖学项目(GUDMAP)”的响应。这是GUDMAP计划(GUDMAP II)的延续，旨在定义下尿路的分子解剖学。我们建议创造一系列新的小鼠品系，使通过这种RFA资助的组织能够识别、分离和遗传操作靶细胞类型，以更好地了解这些细胞及其后代在下尿路发育中的作用。我们预计，这些菌株将使基于抗体的方法能够用于器官解剖的二维和三维成像，用于转录表达分析的细胞分离，以及用于探索下尿路发育和成年器官中细胞关系的细胞命运分析。在AIM1中，我们将利用由NIH资助的KOMP倡议及其欧洲同行EUCOMM创建的广泛的靶向胚胎干细胞(ES)库。这些“敲除第一条件等位基因”将作为二次基因打靶的平台，利用双重组酶介导盒交换的方法。在这方面，新的细胞标记(GPP)和基因组修饰酶(CRE-ERT2)被置于特定基因的控制下，这些基因被选择用于在感兴趣的细胞群体中表达。在目标2中，靶向ES细胞将被用来产生嵌合小鼠，并最终产生携带新标记/修饰等位基因的新小鼠品系。这些毒株将分发给GUDMAP II成员，以方便他们的分析，并分发给NIH资助的小鼠突变资源中心，以便分发给生物医学研究界。

DESCRIPTION (provided by applicant): The proposed research plan is in response to RFA-DK-11-001, "A Genitourinary Anatomy Project (GUDMAP)." This continuation of the GUDMAP initiative (GUDMAP II) aims to define the molecular anatomy of the lower urinary tract. We propose to generate a series of novel mouse strains that will enable groups funded through this RFA to identify, isolate, and genetically manipulate target cell types to better understand the roles of these cells and their descendants in development of the lower urinary tract. We anticipate that the strains will enable antibody-based approaches for 2- and 3-dimensional imaging of organ anatomy, cell isolation for transcriptional analysis of gene expression, and cell fate analysis to explore cell relationships in developing and adult organs of the lower urinary tract. In Aim1, we will make use of the extensive repertoire of targeted embryo stem (ES) cells created by the NIH-funded KOMP initiative and its European counterpart, EUCOMM. These "knock-out first conditional alleles" will serve as a platform for secondary gene-targeting, utilizng a dual recombinase mediate cassette exchange approach. In this, novel cell markers (GPP) and genome modifying enzymes (CRE-ERT2) are placed under the control of specific genes selected for expression in cell populations of interest. In Aim 2, targeted ES cells will be used t generate chimeric mice and ultimately novel mouse strains harboring the new marker/modifier alleles. These strains will be distributed to GUDMAP II members to facilitate their analyses and to NIH-funded Mouse Mutant Resource Centers for distribution to the biomedical research community.