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Updated: Jan 7, 2026
Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes
Published on: May 23, 2025
Genome-wide CRISPR screen reveals an uncharacterized spliceosome regulator as new candidate immunotherapy target
Tong Shao1, Chuanyang Liu1, Jingyu Kuang1
1College of Science National University of Defense Technology Changsha China.
Abstract:
Cancer immune evasion is orchestrated by tumor-intrinsic molecular constraints that remain incompletely defined. Here, we performed an in vivo genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) loss-of-function screen to catalogue gene regulatory determinants of immune evasion in cancer cells. We identify C9ORF50 as a novel splicing regulator whose inhibition profoundly sensitizes cancer to immune surveillance. Integrated multi-omics profiling reveals this intrinsically disordered protein exhibits liquid-liquid phase separation properties and forms nuclear condensates that colocalize with spliceosome components. Genetic ablation correlates with intron retention in multiple spliceosome components and cytoplasmic accumulation of double-stranded RNA, which is associated with type I interferon activation and enhances chemokine-mediated T cell recruitment. As a result, C9ORF50 inhibition amplifies tumor cell immunogenicity, enhancing T cell infiltration in poorly infiltrated tumors. Clinically, elevated C9ORF50 expression correlates with poor survival and diminished lymphoid infiltration across malignancies. Therapeutic targeting of C9ORF50 using RNA interference enhances T cell infiltration and suppresses tumor growth. Our work identifies C9ORF50 as a candidate therapeutic target that modulates RNA splicing and tumor immunity, suggesting splicing regulation as a potential strategy to enhance immunotherapy responses.
Insights
We discovered C9ORF50, a novel splicing regulator, that controls cancer immune evasion. Inhibiting C9ORF50 enhances anti-tumor immunity and T cell infiltration, offering a new therapeutic strategy for cancer immunotherapy.
Area of Science:
- Cancer Biology
- Immunology
- Molecular Biology
Background:
- Cancer immune evasion is a complex process driven by tumor-intrinsic factors.
- Key molecular regulators of immune evasion remain incompletely defined.
- Understanding these regulators is crucial for developing effective cancer immunotherapies.
Purpose of the Study:
- To identify novel gene regulatory determinants of immune evasion in cancer cells.
- To investigate the role of C9ORF50 in regulating cancer immunity.
- To explore C9ORF50 as a potential therapeutic target for enhancing immunotherapy.
Main Methods:
- Performed an in vivo genome-wide CRISPR loss-of-function screen.
- Utilized integrated multi-omics profiling to analyze C9ORF50 function.
- Assessed the impact of C9ORF50 inhibition on tumor immunogenicity and T cell infiltration.
Main Results:
- Identified C9ORF50 as a novel splicing regulator that, when inhibited, sensitizes cancer to immune surveillance.
- C9ORF50 forms nuclear condensates involved in splicing, and its ablation leads to intron retention and cytoplasmic double-stranded RNA accumulation.
- C9ORF50 inhibition enhances type I interferon activation, increases chemokine-mediated T cell recruitment, and improves T cell infiltration in poorly infiltrated tumors.
- Elevated C9ORF50 expression correlates with poor patient survival and reduced lymphoid infiltration across various cancers.
Conclusions:
- C9ORF50 is a critical regulator of RNA splicing and tumor immunity.
- Targeting C9ORF50 enhances cancer immunogenicity and T cell infiltration, suppressing tumor growth.
- Splicing regulation represents a promising strategy to improve immunotherapy responses in cancer patients.
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