The neuroprotective E3-ubiquitin ligase CHIP is linked to healthy aging. Here, we present a protocol using a patient-derived iPSC line with a triplication of the α-synuclein gene to produce gene-edited cells isogenic for CHIP. We describe iPSC differentiation into cortical neurons and their identity validation. We then detail mass spectrometry-based approaches (SWATH-MS) to identify dominant changes in the steady state proteome generated by loss of CHIP function. This protocol can be adapted to other proteins that impact proteostasis in neurons. For complete details on the use and execution of this protocol, please refer to Dias et al. (2021).

• Klíčová slova
• CRISPR, Cell Biology, Cell Differentiation, Cell culture, Mass Spectrometry, Neuroscience, Proteomics, Stem Cells,
• MeSH
• hmotnostní spektrometrie MeSH
• indukované pluripotentní kmenové buňky * MeSH
• lidé MeSH
• neurony MeSH
• proteom genetika   MeSH
• proteomika metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• proteom MeSH