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Updated: Nov 9, 2025
CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions
Published on: November 1, 2024
Generation of GPAM knockout human embryonic stem cell line SYSUe-008-A using CRISPR/Cas9
Chuanbo Sun1, Bing Li2, Miaomiao Yang3
1Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510275, China; Laboratory of Medical Systems Biology, Guangzhou Institute of Pediatrics, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou 510623, China.
Researchers created a glycerol-3-phosphateacyltransferase1 (GPAM) knockout stem cell line to model cerebral palsy. This GPAM knockout cell line can be used to study hypomyelination and develop new therapeutic strategies.
Area of Science:
- Biochemistry
- Genetics
- Neuroscience
Background:
- Glycerol-3-phosphateacyltransferase1 (GPAM) is crucial for lipid biosynthesis.
- GPAM deficiency causes hypomyelination, a hallmark of cerebral palsy.
- Modeling rare genetic diseases like cerebral palsy requires advanced cellular tools.
Purpose of the Study:
- To generate a human embryonic stem cell line lacking functional GPAM.
- To establish a model for studying GPAM-related hypomyelination and cerebral palsy.
- To validate the utility of the knockout cell line for disease modeling.
Main Methods:
- CRISPR/Cas9 gene editing was used to create the GPAM knockout.
- Human embryonic stem cells (SYSUe-008-A) were genetically modified.
- Standard stem cell characterization techniques were employed.
Main Results:
- A GPAM knockout human embryonic stem cell line (SYSUe-008-A) was successfully generated.
- The knockout cell line maintained a normal karyotype.
- Pluripotency markers and differentiation potential were comparable to wild-type cells.
Conclusions:
- The generated GPAM knockout stem cell line serves as a valuable tool for modeling GPAM-related hypomyelination.
- This model facilitates research into the mechanisms of cerebral palsy and the development of targeted therapies.
- The cell line's stability and differentiation capacity support its use in disease modeling studies.
