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Updated: Sep 3, 2025
Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
Published on: June 6, 2020
Efficient Identification of the MYC Regulator with the Use of the CRISPR Library and Context-Matched Database
Yosuke Tanaka1, Hidetaka Kambayashi1, Akiko Yamamoto1,2
1Department of Comprehensive Pathology, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo 113-8510, Japan.
Abstract:
MYC is a major oncogene that plays an important role in cell proliferation in human cancers. Therefore, the mechanism behind MYC regulation is a viable therapeutic target for the treatment of cancer. Comprehensive and efficient screening of MYC regulators is needed, and we had previously established a promoter screening system using fluorescent proteins and the CRISPR library. For the efficient identification of candidate genes, a database was used, for which mRNA expression was correlated with MYC using datasets featuring "Similar" and "Not exactly similar" contexts. INTS14 and ERI2 were identified using datasets featuring the "Similar" context group, and INTS14 and ERI2 were capable of enhancing MYC promoter activity. In further database analysis of human cancers, a higher expression of MYC mRNA was observed in the INTS14 mRNA high-expressing prostate and liver cancers. The knockdown of INTS14 in prostate cell lines resulted in decreased MYC mRNA and protein expression and also induced G0/1 arrest. This study confirmed that CRISPR screening combined with context-matched database screening is effective in identifying genes that regulate the MYC promoter. This method can be applied to other genes and is expected to be useful in identifying the regulators of other proto-oncogenes.
Insights
Researchers identified INTS14 and ERI2 as novel regulators of the MYC oncogene using CRISPR screening and database analysis. Knockdown of INTS14 decreased MYC expression and halted cancer cell proliferation, validating this screening approach for cancer gene discovery.
Area of Science:
- Cancer Biology
- Molecular Oncology
- Gene Regulation
Background:
- The MYC oncogene is a critical driver of cell proliferation in numerous human cancers.
- Understanding MYC regulation is essential for developing targeted cancer therapies.
- Efficient screening methods are needed to identify novel MYC regulators.
Purpose of the Study:
- To develop and validate a comprehensive screening system for identifying MYC regulators.
- To discover novel genes that modulate MYC promoter activity.
- To investigate the role of identified regulators in cancer progression.
Main Methods:
- Established a promoter screening system utilizing fluorescent proteins and CRISPR library.
- Correlated mRNA expression with MYC using context-matched database analysis (Similar and Not exactly similar contexts).
- Validated candidate gene function through promoter activity assays, cancer database analysis, and gene knockdown experiments in prostate cancer cell lines.
Main Results:
- Identified INTS14 and ERI2 as enhancers of MYC promoter activity using the 'Similar' context group.
- Observed higher MYC mRNA expression in prostate and liver cancers with high INTS14 mRNA expression.
- Demonstrated that INTS14 knockdown in prostate cancer cells reduces MYC expression and induces cell cycle arrest at G0/1 phase.
Conclusions:
- CRISPR screening combined with context-matched database analysis is an effective strategy for identifying MYC promoter regulators.
- INTS14 plays a significant role in regulating MYC expression and promoting cancer cell proliferation.
- This methodology can be broadly applied to discover regulators of other proto-oncogenes for cancer therapy development.
