Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics.
STAR Protoc
; 3(2): 101247, 2022 06 17.
Article
em En
| MEDLINE
| ID: mdl-35391935
ABSTRACT
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).
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Células-Tronco Pluripotentes Induzidas
Limite:
Humans
Idioma:
En
Revista:
STAR Protoc
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
Reino Unido