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.
Abnormal development of the lower urinary tract often results in birth defects, whereas in the aging population this system frequently is affected by impaired function and disease. The proposed research aims to generate novel strains of mice that enable the identification, isolation and genetic manipulation of key cell populations from the lower urinary tract. And improved knowledge of the molecular mechanisms underlying urinary tract diseases will, in the future, lead to new treatments in the clinic
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