Related Experiment Video
Updated: Dec 17, 2025
Generation of a RIP1 Knockout U937 Cell Line Using the CRISPR-Cas9 System
Published on: April 11, 2025
Generation of PKD1 mono-allelic and bi-allelic knockout iPS cell lines using CRISPR-Cas9 system
Elena Romano1, Piera Trionfini1, Osele Ciampi1
1Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy.
Abstract:
Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disease, characterised by the development of multiple fluid-filled cysts in the kidneys and other organs. PKD1 and PKD2 are the two major causative genes encoding for polycystin-1 and polycystin-2, respectively. Here, we report the generation of two isogenic induced pluripotent stem cell (iPSC) lines with either heterozygous or compound heterozygous mutations in the PKD1 gene using CRISPR-Cas9 technology. The PKD1+/- and PKD1-/- iPSCs maintain stem cell-like morphology, normal karyotype, pluripotency and differentiation capacity in the three germ layers.
Insights
Researchers created stem cells with mutations in the PKD1 gene, the cause of autosomal dominant polycystic kidney disease (ADPKD). These engineered cells offer a new model for studying ADPKD progression and potential treatments.
Area of Science:
- Genetics and Genomics
- Stem Cell Biology
- Nephrology
Background:
- Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited kidney disorder.
- Mutations in the PKD1 gene are a primary cause of ADPKD, leading to kidney cyst formation.
- Understanding ADPKD pathogenesis requires robust cellular models.
Purpose of the Study:
- To generate isogenic induced pluripotent stem cell (iPSC) lines with specific PKD1 gene mutations.
- To establish a cellular model for studying autosomal dominant polycystic kidney disease (ADPKD).
- To validate the pluripotency and differentiation potential of the generated iPSC lines.
Main Methods:
- CRISPR-Cas9 gene editing technology was employed.
- Two isogenic iPSC lines were generated: one heterozygous (PKD1+/-) and one compound heterozygous (PKD1-/-) for PKD1 mutations.
- Characterization included assessment of stem cell morphology, karyotype, pluripotency markers, and differentiation capacity.
Main Results:
- Isogenic iPSC lines with targeted PKD1 mutations were successfully generated.
- The PKD1+/- and PKD1-/- iPSCs exhibited normal stem cell morphology.
- These engineered iPSCs maintained a normal karyotype and demonstrated pluripotency and differentiation potential across the three germ layers.
Conclusions:
- The study successfully created isogenic iPSC lines carrying PKD1 mutations relevant to ADPKD.
- These novel iPSC lines serve as a valuable tool for investigating ADPKD mechanisms.
- The findings support the utility of these engineered stem cells for future ADPKD research and therapeutic development.
