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Analysis of Nonhomologous End Joining and Homologous Recombination Efficiency in HEK-293T Cells Using GFP-Based Reporter Systems
Published on: February 2, 2024
A PCR-based assay for distinguishing between 293, 293T, and 293E cell lines
Zhilong Wang1, Wenqing Gan1, Qiuyu Chen1
1Key Laboratory of Tropical Translational Medicine of Ministry of Education, International Center for Aging and Cancer, Hainan Medical University, Haikou, 571199, China.
Analytical Biochemistry
|June 16, 2026
Summary
Misidentified human embryonic kidney (HEK) 293 cell sublines risk research reproducibility. A new PCR assay reliably distinguishes 293, 293T, and 293E lines, ensuring experimental accuracy.
Area of Science:
- Cell Biology
- Molecular Biology
- Biotechnology
Background:
- Human embryonic kidney (HEK) 293 cells and their derivatives (293T, 293E) are crucial for biological research and bioproduction.
- Cell line misidentification is a significant issue, compromising experimental reproducibility and data integrity.
Purpose of the Study:
- To develop a reliable method for distinguishing between HEK 293, 293T, and 293E cell lines.
- To provide a cost-effective tool for authenticating these commonly used cell lines.
Main Methods:
- Development of a Polymerase Chain Reaction (PCR)-based assay.
- Validation of the assay using five distinct 293 sublines.
Main Results:
- The PCR assay successfully and accurately identified each of the tested 293 sublines.
- Demonstrated reliability in distinguishing between 293, 293T, and 293E cell lines.
Conclusions:
- The developed PCR assay is a simple and effective tool for authenticating HEK 293 sublines.
- Ensures the correct cell line is utilized in research and bioproduction, enhancing data reliability.

