From RNAi to single-cell CRISPR: Evolving functional screening in aging and cancer

Abstract

Aging and cancer represent divergent fates of a common stress response. Senescence initially restrains malignancy through stable cell-cycle arrest and immune clearance, yet persistence of senescent cells and their secretory phenotype (SASP) paradoxically drives tumor progression, therapeutic resistance, and immune evasion via chronic inflammation and metabolic reprogramming. Functional genomic screening has evolved from bulk RNA interference to precision CRISPR modalities, including knockout, interference, activation, and base editing, and onward to single-cell and in vivo platforms. These technological advances have transformed our capacity to dissect cellular state transitions and network dependencies with unprecedented resolution. Here, we trace this technological evolution and synthesize its impact on aging and cancer research, with emphasis on chromatin regulation, metabolic rewiring, the SASP, nucleocytoplasmic transport, therapeutic response, and the tumor microenvironment. We emphasize that the current evidence derives predominantly from preclinical functional screens and computational predictions. We propose shifting from gene-centric discovery toward context-dependent network analysis. Integrating precise editing, in vivo screening, single-cell multi-omics, and emerging AI-assisted design may provide information and a design basis for future combined strategies that simultaneously target vulnerabilities in senescent cells and malignant populations.

Keywords

CRISPR screening / senescence / cancer / functional genomics

1 Declarations

1.1 Acknowledgements

The authors would like to thank all the researchers whose work is included in this review. We acknowledge the use of AI-assisted tools, including DeepSeek, for language editing and polishing of the manuscript. All authors have full control over the content of the manuscript, have thoroughly reviewed and revised all parts of the manuscript, and take full responsibility for all content of the manuscript.

2 Author contributions

Fan BT: Writing-original draft.

Wu AW: Investigation, supervision.

Pan X, Wang H: Conceptualization, writing-review & editing.

3 Conflicts of interest

The authors declare no competing interests.

4 Ethical approval

Not applicable.

5 Consent to participate

Not applicable.

6 Consent for publication

Not applicable.

7 Availability of data and materials

Not applicable.

Funding

This work was supported by the National Key R&D Program of China (2021YFA1102800 to H.W.) and by National Natural Science Foundation of China (82301746 to A.W.); “Pioneer” and “Leading Goose” R&D Program of Zhejiang (No.2026C02A1147); the Hangzhou Youth Innovation Team Project (No. TD2023020 to A.W.); the HZNU scientific research and innovation team project (No. TD2025002 to H.W).

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