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Updated: Jan 23, 2026
Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
Published on: June 6, 2020
A large-scale CRISPR screen and identification of essential genes in cellular senescence bypass
Xuehui Liu1,2, Lei Wei1, Qiongye Dong1,3
1MOE Key Laboratory of Bioinformatics; Bioinformatics Division and Center for Synthetic and Systems Biology, Beijing National Research Center for Information Science and Technology, Department of Automation, Tsinghua University, Beijing 100084, China.
Abstract:
Cellular senescence is an important mechanism of autonomous tumor suppression, while its consequence such as the senescence-associated secretory phenotype (SASP) may drive tumorigenesis and age-related diseases. Therefore, controlling the cell fate optimally when encountering senescence stress is helpful for anti-cancer or anti-aging treatments. To identify genes essential for senescence establishment or maintenance, we carried out a CRISPR-based screen with a deliberately designed single-guide RNA (sgRNA) library. The library comprised of about 12,000 kinds of sgRNAs targeting 1378 senescence-associated genes selected by integrating the information of literature mining, protein-protein interaction network, and differential gene expression. We successfully detected a dozen gene deficiencies potentially causing senescence bypass, and their phenotypes were further validated with a high true positive rate. RNA-seq analysis showed distinct transcriptome patterns of these bypass cells. Interestingly, in the bypass cells, the expression of SASP genes was maintained or elevated with CHEK2, HAS1, or MDK deficiency; but neutralized with MTOR, CRISPLD2, or MORF4L1 deficiency. Pathways of some age-related neurodegenerative disorders were also downregulated with MTOR, CRISPLD2, or MORF4L1 deficiency. The results demonstrated that disturbing these genes could lead to distinct cell fates as a consequence of senescence bypass, suggesting that they may play essential roles in cellular senescence.
Insights
Identifying genes that regulate cellular senescence is crucial for cancer and aging therapies. This study used CRISPR screening to find genes controlling senescence bypass, revealing key regulators like MTOR and CHEK2 that influence cell fate and disease pathways.
Area of Science:
- Cellular and Molecular Biology
- Genetics and Genomics
- Cancer Research
Background:
- Cellular senescence is a key tumor suppressor mechanism.
- The senescence-associated secretory phenotype (SASP) can promote cancer and age-related diseases.
- Controlling cell fate during senescence is vital for anti-cancer and anti-aging strategies.
Purpose of the Study:
- To identify genes critical for establishing or maintaining cellular senescence.
- To investigate the role of specific genes in senescence bypass.
- To understand how gene deficiencies impact cell fate and associated pathways.
Main Methods:
- Utilized a CRISPR-based screen with a custom sgRNA library targeting 1378 senescence-associated genes.
- Integrated literature mining, protein-protein interaction networks, and differential gene expression for gene selection.
- Validated identified gene deficiencies and analyzed transcriptome patterns using RNA-seq.
Main Results:
- Identified several gene deficiencies that lead to senescence bypass with high accuracy.
- Observed distinct transcriptome profiles in bypass cells.
- Found that CHEK2, HAS1, or MDK deficiency maintained/elevated SASP, while MTOR, CRISPLD2, or MORF4L1 deficiency neutralized it.
- Noted downregulation of age-related neurodegenerative disorder pathways with MTOR, CRISPLD2, or MORF4L1 deficiency.
Conclusions:
- Disturbing specific genes identified in this screen can lead to distinct cell fates following senescence bypass.
- These genes play essential roles in regulating cellular senescence.
- Findings offer potential therapeutic targets for cancer and age-related diseases.
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