PI: Zbyněk Kozmik Conditional Mice Mutants as Tools for Studying Genetic Disorders and Eye Physiology

The mouse model represents an attractive alternative for studying the development and pathogenesis of the human eye. Currently, a systematic targeted mutagenesis of mice genes in the embryonic stem cells is being done (EUCOMM and KOMP consortia). Shortly, the conditional mutants for the majority of the genes will be available for phenotypic analysis. Their utility is, to a great extent, dependent on the availability of suitable strains of mice with the tissue or time limited activity of the Cre recombinase. For each area of interest, e.g. eye, brain, liver, it is necessary to have a panel of the Cre-transgenic mouse strains which enable a specific deletion of the given gene and a detailed study of its function in the context of the whole organism. The size of the Cre panel which it is necessary to create, is dependent primarily on the complexity of an organ (number of cellular types, development stages), but also by the current availability of Cre lines for the given organ.

For instance, the Cre expression in the very early embryonic retina, in certain types of retinal cells of an adult mouse and in the lens and the cornea of an adult mouse is not available for the functional genomics of the eye so far.

The project will be focused on the systematic preparation of new transgenic Cre lines with defined expressions in the eye tissues and the consecutive usage of the Cre lines for the systematic analysis of the conditional mutants in genes coding the transcriptional factors and components of the Wnt signaling pathway.

The new transgenic mice lines with the tissue and time limited activity of the Cre recombinase will be prepared in the presented project. Their utility will be verified by the deletion of genes which have the known pathogenesis. Then the suitable Cre lines will be used for the systematic gene deletions and the study of their function in eye tissues. The project will contribute to our understanding of eye diseases. Goal: 1/ To prepare the transgenic Cre constructs with the BAC recombineering technology and with their help to create the Cre lines for the functional genomic of the early retina and adult cornea, 2/ to prepare the transgenic constructs with the pharmacologically controlled form of the Cre recombinase (Cre-ERTM) and to create the Cre lines with time conditioned activity of the Cre recombinase, 3/ to map the function of the genes encoding transcription factors and components of the Wnt signaling pathway in the eye tissues.