MAGEL2 is one of the largest members of the melanoma antigen (MAGE) protein family, encoded within the critical 15q11-q13 region of chromosome 15. This region is crucial in Prader-Willi Syndrome (PWS), a multisystem neurodevelopmental disorder. Genes within this PWS-critical region are maternally imprinted and expressed solely from the paternal allele; their loss of expression results in PWS. Besides the deletion of the whole region, the truncating mutations in the gene MAGEL2 lead to a PWS-like syndrome, referred to as Schaaf-Yang Syndrome (SYS). Several molecular characteristics underlying PWS and SYS have been elucidated in the last 10 years, however, the exact mechanism of their development and targeted therapy for patients still await discovery. Recent data suggested that MAGEL2 regulates retromer-dependent intracellular recycling of proteins and endocrine function of hypothalamus; but the physiological function of MAGEL2 and the pathological consequences of its absence are still not clear. Recently a polyclonal antibody against the mouse Magel2 protein was developed, which will, for the first time, allow determination of MAGEL2 localization in situ and determine binding partners in physiologically relevant tissues where MAGEL2 is expressed. The aim of my master’s thesis was to perform the preliminary analysis and optimization for the immunoprecipitation studies. We examined the tissue expression profile of MAGEL2, and its known binding partners using publicly available databases and optimized the immunoprecipitation protocol. This optimized approach will be applied in future studies to identify physiologically relevant MAGEL2 interactors and its molecular roles in mouse tissues.
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