Novel CRISPR-Cas9 iPSC knockouts for PCCA and PCCB genes: advancing propionic acidemia research
Propionic acidemia (PA) is a rare autosomal recessive metabolic disorder caused by mutations in the PCCA and PCCB genes, which encode subunits of the mitochondrial enzyme propionyl-CoA carboxylase (PCC). This enzyme deficiency leads to the accumulation of toxic metabolites, resulting in severe metabolic dysfunction. To create ideal in vitro disease models of PA with isogenic controls and provide a robust platform for therapeutic research, we generated two induced pluripotent stem cell (iPSC) lines with knockout (KO) mutations in the PCCA and PCCB genes using CRISPR-Cas9 gene editing in a healthy control iPSC line. The KO iPS cells were successfully established and characterized, confirming the presence of frameshift insertions and deletions in each target gene, as well as the loss of the corresponding transcript, protein expression, and activity. Additionally, the generated iPSC lines exhibit hallmark characteristics of pluripotency, including the potential to differentiate into all three germ layers. Our PCCA and PCCB KO iPSC models provide a valuable tool for studying the molecular mechanisms underlying PA and hold potential for advancing new therapeutic approaches.
Authors
- Lourdes R. Desviat (ORCID: https://orcid.org/0000-0003-0492-3323)
- Eva Richard (ORCID: https://orcid.org/0000-0001-9711-1417)
- Mar Álvarez (ORCID: https://orcid.org/0000-0002-6086-6557)
- Emilio M. García-Tenorio
- Mónica Gallego-Bonhomme
Institutions
- Instituto de Salud Carlos III (ES)
- Hospital La Paz Institute for Health Research (ES)
- Centre for Biomedical Network Research on Rare Diseases (ES)
- Centro de Investigación Biomédica en Red (ES)
- Centro de Biología Molecular Severo Ochoa (ES)
- Universidad Autónoma de Madrid (ES)
Publication Details
- Journal
- Human Cell
- Published
- 2025-03-05
- DOI
- https://doi.org/10.1007/s13577-025-01193-z
- Citations
- 1
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 0.81
Funders
- Fundación Ramón Areces
- Centro de Biología Molecular Severo Ochoa
- Propionic Acidemia Foundation
- Universidad Autónoma de Madrid
- Ministerio de Ciencia e Innovación