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Updated: Dec 30, 2025
Generation of a RIP1 Knockout U937 Cell Line Using the CRISPR-Cas9 System
Published on: April 11, 2025
Generation of a DAPK1 knockout first (conditional ready) human embryonic stem cell line (ZSSYe001-A) by CRISPR-Cas9
Cancan Xu1, Zhuowei Zhou2, Chang Liu2
1Institute of Physical Science and Information Technology, Anhui University, Hefei 230601, China; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
Abstract:
Death-associated protein kinase 1 (DAPK1) is a Ca2+/calmodulin regulated Ser/Thr kinase involved in various cellular processes including cell death, autophagy and inflammation. Its dysregulation has been linked to tumour metastasis, anti-viral responses, Alzheimer's disease and other neurological disorders. To further investigate the role of DAPK1 in these processes, we generated a DAPK1 knockout first (conditional ready) human embryonic stem (hES) cell line in which the endogenous DAPK1 can be easily restored with expression of FLPe. This cell line provides an ideal model to study the role of DAPK1 in human development and various pathologies related to DAPK1 dysregulation in vitro.
Insights
Researchers created a new human stem cell model lacking Death-associated protein kinase 1 (DAPK1). This model allows easy DAPK1 restoration, aiding study of its role in development and diseases like cancer and Alzheimer's.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Death-associated protein kinase 1 (DAPK1) is a calcium/calmodulin-regulated serine/threonine kinase.
- DAPK1 plays roles in crucial cellular processes, including cell death, autophagy, and inflammation.
- Dysregulation of DAPK1 is implicated in tumor metastasis, antiviral responses, Alzheimer's disease, and other neurological disorders.
Purpose of the Study:
- To develop a versatile human cellular model for investigating DAPK1 functions.
- To facilitate the study of DAPK1's role in human development and disease pathogenesis.
- To enable precise control over DAPK1 expression for in vitro studies.
Main Methods:
- Generation of a DAPK1 knockout first (conditional ready) human embryonic stem (hES) cell line.
- Utilizing FLPe recombinase system for inducible restoration of endogenous DAPK1 expression.
- Employing CRISPR/Cas9 or similar gene-editing technologies for generating the knockout.
Main Results:
- Successfully created a conditional ready hES cell line with DAPK1 specifically knocked out.
- Established a system for efficient and controllable re-expression of DAPK1 in the knockout cells.
- Validated the functionality of the generated hES cell line as a model system.
Conclusions:
- The developed DAPK1 knockout hES cell line is a powerful tool for studying DAPK1 biology.
- This model system is suitable for investigating DAPK1's involvement in human development and disease.
- Facilitates in vitro research into pathologies associated with DAPK1 dysregulation.
