Generation of induced pluripotent stem cell lines carrying monoallelic (UCSFi001-A-60) or biallelic (UCSFi001-A-61; UCSFi001-A-62) frameshift variants in CACNA1A using CRISPR/Cas9

Marina P Hommersom; Chantal Bijnagte-Schoenmaker; Silvia Albert; Bart P C van de Warrenburg; Nael Nadif Kasri; Hans van Bokhoven

doi:10.1016/j.scr.2022.102730

Generation of induced pluripotent stem cell lines carrying monoallelic (UCSFi001-A-60) or biallelic (UCSFi001-A-61; UCSFi001-A-62) frameshift variants in CACNA1A using CRISPR/Cas9

Stem Cell Res. 2022 May:61:102730. doi: 10.1016/j.scr.2022.102730. Epub 2022 Feb 26.

Authors

Marina P Hommersom¹, Chantal Bijnagte-Schoenmaker¹, Silvia Albert¹, Bart P C van de Warrenburg², Nael Nadif Kasri³, Hans van Bokhoven⁴

Affiliations

¹ Department of Human Genetics, Donders Institute for Brain, Cognition, and Behaviour, Radboud University Medical Center, 6500 HB Nijmegen, the Netherlands.
² Department of Neurology, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6500 HB Nijmegen, the Netherlands.
³ Department of Human Genetics, Donders Institute for Brain, Cognition, and Behaviour, Radboud University Medical Center, 6500 HB Nijmegen, the Netherlands; Department of Cognitive Neuroscience, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6500 HB Nijmegen, Netherlands.
⁴ Department of Human Genetics, Donders Institute for Brain, Cognition, and Behaviour, Radboud University Medical Center, 6500 HB Nijmegen, the Netherlands; Department of Cognitive Neuroscience, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6500 HB Nijmegen, Netherlands. Electronic address: [email protected].

PMID: 35286975
DOI: 10.1016/j.scr.2022.102730

Abstract

CACNA1A encodes a P/Q-type voltage-gated calcium channel. Heterozygous loss-of-function variants in this gene have been associated with episodic ataxia type 2. In this study, we used CRISPR/Cas9 to generate isogenic human induced pluripotent stem cell lines with a gene-dosage dependent deficiency of CACNA1A. We obtained one clone with monoallelic (UCSFi001-A-60) and two clones with biallelic (UCSFi001-A-61; UCSFi001-A-62) frameshift variants in CACNA1A. All three lines showed expression of pluripotency markers and a normal karyotype.

Keywords: CACNA1A; ataxia; neuron.

MeSH terms

CRISPR-Cas Systems / genetics
Calcium Channels / metabolism
Cells, Cultured
Frameshift Mutation
Humans
Induced Pluripotent Stem Cells* / metabolism

Substances

CACNA1A protein, human
Calcium Channels