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Participant 4

Georg-August-University Göttingen

Partner 4: University Göttingen, Biochemistry II, AG Landgrebe, D-37073, Göttingen (Team leader/Responsible Scientist: Dr. Torben Lübke); Key skills: Transcriptome analysis

Laboratory background and technical information: The department of Biochemistry 2 is part of the medical faculty of the University of Göttingen and is engaged in teaching programs for students of medicine and biology. It has all facilities necessary for mouse experiments. The majority of the projects is focussed on the biogenesis of lysososomes in mammalian cells. The institute has a long standing interest in genetic defects that result in lysosomal storage disorders. Another important research branch of the institute is the biochemical and molecular analysis of congenital disorders of glycosylation (CDG-Syndromes) using patient cells and mouse mutants as models. Since two years, the department has a microarray facility to perform transcriptome analysis experiments.

Capacity to contribute to the project: We have established a central microarray facility of the faculty of medicine at our institute and begun to analyse gene expression in alpha mannosidase deficient animals, wildtype controls and alpha mannosidase deficient animals treated with enzyme replacement therapy (ERT) in collaboration with partner 1. To perform transcriptome analysis experiments, we use custom made two-colour cDNA microarrays featuring about 20.000 mouse genes as well as commercial two-colour oligomicroarrays with 44.000 features representing 35.000 mouse genes (Agilent). The group is experienced in the usage of microarrays since 5 years and specialised in statistical microarray data analysis as well as functional gene annotations using applied bioinformatics and transcriptome analysis follow-up experiments.

Members involved: • Dipl. biol. NN: One PhD student to perform tissue and RNA preparations from alpha mannosidase-deficient mice with and without treatment as well as wild-type controls. Different tissues and treatment durations will be analysed using high-density two-colour cDNA- and oligo-microarrays. Follow-up experiments to validate the microarray results as well as to obtain conclusive biochemical and cell biological findings will be performed (work packages 1, 2 and 5) (36 pm). • One diploma student to perform biochemical and cell biological follow up experiments based on the transcriptome findings (6 pm). The costs to perform the microarray experiments described in workpackage 5 will amount to €50.000. These costs should be covered by HUE-MAN. The costs for follow-up expriments can be met by the basic funds of the institute.

Other contributing members of the laboratory: Dr. Torben Lübke, group leader: supervision of the project, microarray data analysis and annotation, design of followup experiments (18 pm).

TOTAL EXPECTED EFFORT: 60 pm

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