Large Scale Screen for Genes Controlling Mammalian Embryogenesis

Bernhard Herrmann
Max-Planck-Institut für Immunbiologie
Dept. Developmental Biology
Stuebeweg 51
79108 Freiburg
telephone: +49 761 5108 582
fax: +49 761 5108 569
prestype: Platform
presenter: Bernhard Herrmann

Lorenz Neidhardt, Stephan Gasca, Karin Wertz, Andrea Schepler, Franz Obermayr, Alexander Aulehla, Susanne Worpenberg and Bernhard G. Herrmann

The molecular analysis of cell differentiation, tissue patterning and organ development requires the isolation of genes involved in these processes. Large scale EST and genome sequencing have been employed by many laboratories in order to identify all genes encoded by the human genome. However, sequence information alone does not reveal the tissue type, process or organ in which a particular gene is acting.

We have developed a high-throughput strategy for determining the expression of genes in 9.5 day mouse embryos on a large scale. Genes are selected from cDNA libraries or cDNA arrays and assayed for their expression patterns by multiple whole mount in situ hybridisation analyses, thus providing expression data at high resolution. A genome set of 40,000 genes could be assayed in 20 person years (i.e. 5 persons, 4 years). Gene expression and sequence information together represent essential information about the cell type and process a gene is acting in, and often also about the biochemical function. This information can be utilized to develop strategies for further functional analyses, and to identify potential target genes for pharmaceutical drugs. In addition, chromosomal mapping of the genes identifies candidates for known human disorders and mouse mutations.

In a pilot screen, appr. 800 specifically expressed genes have been identified. Of 607 clones sequenced, about 43% represent known genes and about 44% novel genes or ESTs. The remaining cDNAs represent mouse orthologues of known genes or new members of known gene families.

Appr. 25% of the known genes isolated are involved in transcriptional regulation, such as Hox genes, Tbx genes, Msx, Pax9. Appr. 16% of the known genes isolated represent mouse orthologues of human disorders. Three mouse genes have been isolated for which a classical mutant locus is known (Brachyury, pudgy, vibrator). This suggests that a similar proportion of mouse and human mutations is represented among the novel genes and ESTs identified. A data base providing expression, sequence and homology search data is being built up at the Resource Center (RZPD). This project will have a strong impact on several fields of biomedical research.

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