Related Experiment Video
Updated: Sep 3, 2026
Pooled CRISPR-Based Genetic Screens in Mammalian Cells
Published on: September 4, 2019
Genome-scale CRISPR-Cas9 knockout screening in nasopharyngeal carcinoma for radiosensitive and radioresistant genes
Ziyan Zhou1, Gang Chen2, Mingjun Shen1
1Department of Radiation Oncology, the First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi, China; Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor (Guangxi Medical University), Ministry of Education, Nanning, 530021, Guangxi, China; Guangxi Key Laboratory of Immunology and Metabolism for Liver Diseases, Nanning, 530021, Guangxi, China.
Background:
Genome-scale CRISPR-Cas9 knockout screening may provide new insights into the mechanism underlying clinical radioresistance in nasopharyngeal carcinoma (NPC), which is remain largely unknown. Our objective was to screen the functional genes associated with radiosensitivity and radioresistance in NPC, laying a foundation for further research on its functional mechanismand.
Methods:
CRISPR-Cas9 library lentivirus screening in radiation-treated NPC cells was combined with second-generation sequence technology to identify functional genes, which were further validated in radioresistant NPC cells and patient tissues.
Results:
Eleven radiosensitive and radioresistant genes were screened. Among these genes, the expression of FBLN5, FAM3C, MUS81, and DNAJC17 were significantly lower and TOMM20, CDKN2AIP, SNX22, and SP1 were higher in the radioresistant NPC cells (C666-1R, 5-8FR) (p < 0.05). CALD1 was highly expressed in C666-1R. Furthermore, we found knockout of FBLN5, FAM3C, MUS81 and DNAJC17 promoted the proliferation of NPC cells, while CDKN2AIP and SP1 had the opposed results (p < 0.05). This result was verified in NPC patient tissues. Meanwhile, KEGG analysis showed that the Fanconi anemia pathway and the TGF-β signaling pathway possibly contributed to radiosensitivity or radioresistance in NPC.
Conclusions:
Nine genes involved in the radiosensitivity or radioresistance of NPC: four genes for radiosensitivity (FBLN5, FAM3C, MUS81, and DNAJC17), two genes for radioresistance (CDKN2AIP, SP1), two potential radioresistant genes (TOMM20, SNX22), and a potential radiosensitive gene (CALD1). Genome-scale CRISPR-Cas9 knockout screening for radiosensitive and radioresistant genes in NPC may provide new insights into the mechanisms underlying clinical radioresistance to improve the efficacy of radiotherapy for NPC.
Insights
This study used CRISPR-Cas9 screening to identify genes affecting nasopharyngeal carcinoma (NPC) radiosensitivity and radioresistance. Key genes like FBLN5 and CDKN2AIP were identified, offering new targets for improving radiotherapy efficacy in NPC patients.
Area of Science:
- Oncology
- Genetics
- Molecular Biology
Background:
- Nasopharyngeal carcinoma (NPC) radioresistance mechanisms are poorly understood.
- Genome-scale CRISPR-Cas9 screening offers a powerful tool to uncover novel insights.
- Identifying genes influencing radiosensitivity is crucial for improving NPC treatment.
Purpose of the Study:
- To screen for functional genes associated with radiosensitivity and radioresistance in NPC.
- To lay the foundation for understanding the molecular mechanisms of NPC radioresistance.
- To identify potential therapeutic targets for enhancing radiotherapy efficacy.
Main Methods:
- Utilized CRISPR-Cas9 library lentivirus screening in radiation-treated NPC cells.
- Employed second-generation sequencing to identify functional genes.
- Validated identified genes in radioresistant NPC cells and patient tissues.
Main Results:
- Screened eleven radiosensitive and radioresistant genes.
- Identified differential expression of FBLN5, FAM3C, MUS81, DNAJC17, TOMM20, CDKN2AIP, SNX22, and SP1 in radioresistant NPC cells.
- Knockout of FBLN5, FAM3C, MUS81, and DNAJC17 promoted NPC cell proliferation, while CDKN2AIP and SP1 showed opposite effects.
- Fanconi anemia and TGF-β signaling pathways implicated in NPC radiosensitivity/resistance.
Conclusions:
- Identified nine genes involved in NPC radiosensitivity or radioresistance.
- Four genes (FBLN5, FAM3C, MUS81, DNAJC17) linked to radiosensitivity.
- Two genes (CDKN2AIP, SP1) linked to radioresistance.
- Potential therapeutic targets identified to improve radiotherapy outcomes in NPC.
Related Concept Videos
02:46Genetic Screens
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...
01:28CRISPR/Cas9 Genome Editing