Generation of homozygous PRKN, PINK1 and double PINK1/PRKN knockout cell lines from healthy induced pluripotent stem cells using CRISPR/Cas9 editing

Carol X-Q Chen; Zhipeng You; Narges Abdian; Julien Sirois; Irina Shlaifer; Mahdieh Tabatabaei; Marie-Noëlle Boivin; Lydiane Gaborieau; Jason Karamchandani; Lenore K Beitel; Edward A Fon; Thomas M Durcan

doi:10.1016/j.scr.2022.102806

Generation of homozygous PRKN, PINK1 and double PINK1/PRKN knockout cell lines from healthy induced pluripotent stem cells using CRISPR/Cas9 editing

Stem Cell Res. 2022 Jul:62:102806. doi: 10.1016/j.scr.2022.102806. Epub 2022 May 4.

Affiliations

¹ The Neuro's Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada.
² C-BIG Repository (C-BIG), Montreal Neurological Institute-Hospital, McGill University, Montreal, QC H3A 2B4, Canada.
³ McGill Parkinson Program and Neurodegenerative Diseases Group, Montreal Neurological Institute, Department of Neurology and Neurosurgery, Montreal, QC H3A 2B4, Canada.
⁴ The Neuro's Early Drug Discovery Unit (EDDU), McGill University, 3801 University Street, Montreal, QC H3A 2B4, Canada. Electronic address: [email protected].

PMID: 35561458
DOI: 10.1016/j.scr.2022.102806

Abstract

Autosomal recessive mutations in either PRKN or PINK1 are associated with early-onset Parkinson's disease. The corresponding proteins, PRKN, an E3 ubiquitin ligase, and the mitochondrial serine/threonine-protein kinase PINK1 play a role in mitochondrial quality control. Using CRISPR/CAS9 technology we generated three human iPSC lines from the well characterized AIW002-02 control line. These isogenic iPSCs contain homozygous knockouts of PRKN (PRKN-KO, CBIGi001-A-1), PINK1 (PINK1-KO, CBIGi001-A-2) or both PINK1 and PRKN (PINK1-KO/PRKN-KO, CBIGi001-A-3). The knockout lines display normal karyotypes, express pluripotency markers and upon differentiation into relevant brain cells or midbrain organoids may be valuable tools to model Parkinson's disease.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics
Cell Line
Humans
Induced Pluripotent Stem Cells* / metabolism
Mitophagy / genetics
Parkinson Disease* / genetics
Protein Kinases / genetics
Ubiquitin-Protein Ligases / genetics
Ubiquitin-Protein Ligases / metabolism

Substances

Ubiquitin-Protein Ligases
Protein Kinases

Grants and funding

PJT – 169095/CIHR/Canada