Related Experiment Video
Updated: Aug 12, 2025
CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions
Published on: November 1, 2024
Establishment of a PPP2CA homozygous knockout human embryonic stem cell line via CRISPR/Cas9 system
Tongbin Ding1, Zhe Gao1, Hang Liu1
1Department of Cardiology, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou 450014, China.
Abstract:
Protein phosphatase 2A (PPP2CA) is one of the four main Ser/Thr phosphatase enzymes, which involved in the negative control of cell growth and division. PPP2CA is the main protein phosphatase in the heart, which regulates Ca (2+) through a series of ion channels and transporters. In this study, we generated a PPP2CA homozygous knockout human embryonic stem cell line WAe009-A-25 based on the transient expression CRISPR/Cas9 system to investigate functional effect of PP1 deficiency. This cell line has multidirectional differentiation potential, normal karyotypic and trilineage differentiation potential in vivo.
Insights
Researchers created a knockout stem cell line lacking Protein Phosphatase 2A (PPP2CA) to study its role in cell growth and heart function. This new cell line maintains normal karyotype and differentiation potential for further research.
Area of Science:
- Biochemistry
- Molecular Biology
- Stem Cell Biology
Background:
- Protein Phosphatase 2A (PPP2CA) is a key Ser/Thr phosphatase regulating cell growth and division.
- PPP2CA plays a critical role in cardiac function, controlling calcium (Ca2+) regulation via ion channels and transporters.
Purpose of the Study:
- To generate a homozygous knockout human embryonic stem cell line for Protein Phosphatase 2A (PPP2CA).
- To investigate the functional consequences of PPP2CA deficiency using CRISPR/Cas9 technology.
Main Methods:
- Utilized CRISPR/Cas9 gene editing technology for transient expression.
- Generated a homozygous knockout human embryonic stem cell line (WAe009-A-25) for PPP2CA.
Main Results:
- Successfully created a PPP2CA homozygous knockout human embryonic stem cell line.
- The generated cell line (WAe009-A-25) exhibits normal karyotypic stability.
- The cell line demonstrates trilineage differentiation potential in vivo, indicating pluripotency is maintained.
Conclusions:
- The developed PPP2CA knockout stem cell line is a valuable tool for studying the phosphatase's function.
- This cell line allows for in-depth investigation into the role of PPP2CA in cellular processes, particularly in cardiac function and development.
