Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics

Catarina Dias; Erisa Nita; Jakub Faktor; Lenka Hernychova; Tilo Kunath; Kathryn L. Ball

doi:10.1016/j.xpro.2022.101247

Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics

Catarina Dias, Erisa Nita, Jakub Faktor, Lenka Hernychova, Tilo Kunath, Kathryn L. Ball^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract / Description of output

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).

Original language	English
Article number	101247
Pages (from-to)	101247
Number of pages	33
Journal	STAR Protocols
Volume	3
Issue number	2
Early online date	2 Apr 2022
DOIs	https://doi.org/10.1016/j.xpro.2022.101247
Publication status	Published - 17 Jun 2022

Keywords / Materials (for Non-textual outputs)

cell biology
cell culture
cell differentiation
CRISPR
mass spectrometry
neuroscience
proteomics
stem cells

Access to Document

10.1016/j.xpro.2022.101247Licence: Creative Commons: Attribution-NonCommercial-NoDerivatives (CC BY-NC-ND)

BallEtalSP2022GenerationOfACHIPIsogenicHumanFinal published version, 6.2 MBLicence: Creative Commons: Attribution-NonCommercial-NoDerivatives (CC BY-NC-ND)

Generating Biologics to Modulate IFITM1 in Cancer and Infection
Ball, K. & Haas, J.
UK-based charities
1/09/17 → 31/03/22
Project: Research
Using iPSC-derived neurons to study homeostatic control of aSyn by the E3-ubiquitin ligase and co-chaperone CHIP
Ball, K.
UK-based charities
1/02/16 → 31/01/19
Project: Research

Cite this

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title = "Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics",

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).",

keywords = "cell biology, cell culture, cell differentiation, CRISPR, mass spectrometry, neuroscience, proteomics, stem cells",

author = "Catarina Dias and Erisa Nita and Jakub Faktor and Lenka Hernychova and Tilo Kunath and Ball, {Kathryn L.}",

year = "2022",

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doi = "10.1016/j.xpro.2022.101247",

language = "English",

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TY - JOUR

T1 - Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics

AU - Dias, Catarina

AU - Nita, Erisa

AU - Faktor, Jakub

AU - Hernychova, Lenka

AU - Kunath, Tilo

AU - Ball, Kathryn L.

PY - 2022/6/17

Y1 - 2022/6/17

N2 - 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).

AB - 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).

KW - cell biology

KW - cell culture

KW - cell differentiation

KW - CRISPR

KW - mass spectrometry

KW - neuroscience

KW - proteomics

KW - stem cells

U2 - 10.1016/j.xpro.2022.101247

DO - 10.1016/j.xpro.2022.101247

M3 - Article

C2 - 35391935

AN - SCOPUS:85127487622

SN - 2666-1667

VL - 3

SP - 101247

JO - STAR Protocols

JF - STAR Protocols

IS - 2

M1 - 101247

ER -

Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics

Abstract / Description of output

Keywords / Materials (for Non-textual outputs)

Access to Document

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Projects

Generating Biologics to Modulate IFITM1 in Cancer and Infection

Using iPSC-derived neurons to study homeostatic control of aSyn by the E3-ubiquitin ligase and co-chaperone CHIP

Cite this