Optogenetic induction of alpha-synuclein aggregation in human dopaminergic neurons to model Parkinson’s disease pathology

Abstract

Summary: Alpha-synuclein (α-syn) aggregation is a principal factor in Parkinson’s disease (PD) onset. Here, we present a protocol for optogenetic induction of α-syn aggregation in human midbrain dopaminergic (mDA) neurons, facilitating a detailed PD pathology study. We describe steps for nucleofection of the opto-α-syn construct, single colony selection and validation, alongside mDA neuron differentiation and rapid induction of toxic α-syn aggregates via blue light. This establishes a potent human induced pluripotent-stem-cell-based platform for PD drug testing and validation.For complete details on the use and execution of this protocol, please refer to Kim et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Stem Cells
• Cell Differentiation
• Organoids