Functional Cloning of Genes Regulating Apoptosis in Neuronal Cells

Here, we describe the use of a functional cloning approach, based on the screen of a genomewide short hairpin RNA (shRNA) library, to identify novel genes regulating apoptosis in neuronal cells. Apoptosis is induced by doxorubicin and is detected with a fluorometric caspase 3 assay. Moreover, we describe also the screen of the library to identify genes regulating the processing of the β-amyloid (Aβ) precursor protein (APP), the protein associated with the pathogenesis of Alzheimer’s disease. The levels of the peptide Aβ, produced by the APP processing, are detected with ELISA based on the innovative Delfia method developed by PerkinElmer.