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RNA exonuclease REXO4 resolves m6A-marked R-loops and suppresses anti-tumor immunity [RNAseq in REXO4-dTAG HeLa cells]

ABSTRACT: To determine if REXO4 depletion directly induces R-loop accumulation independent of transcription, we generated a HeLa cell line expressing endogenous REXO4 tagged with an N-terminal HA-FKBP12/F36V degron (REXO4-dTAG) via microhomology-mediated end joining (MMEJ)-based CRISPR/Cas9. Acute REXO4 degradation, achieved by a 4-hr treatment with dTAG-13 (dTAG), was then analyzed by RNA-seq to assess transcriptomic changes

ORGANISM(S): Homo sapiens

PROVIDER: GSE313849 | GEO | 2026/05/28

REPOSITORIES: GEO

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RNA exonuclease REXO4 resolves m6A-marked R-loops and suppresses anti-tumor immunity [RNAseq in REXO4-dTAG HeLa cells]

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RNA exonuclease REXO4 resolves m6A-marked R-loops and suppresses anti-tumor immunity [RNAseq]

Project description:Transcription blockage frequently occurs in tumor cells, and aberrant R-loop forming during this process drives genome instability. However, the regulation of R-loop homeostasis and its dysregulation in tumorigenesis are still under investigation. Here, we report that the RNA exonuclease REXO4 preserves genome stability by reducing excessive R-loops. Mechanistically, REXO4 resolves R-loops by 3’-5’ exonucleolytic cleavage of the RNA strand on RNA-DNA hybrids, where an accessible end is created by RNA endonuclease and N6-methyladenosine (m6A) modification on RNA moieties promotes REXO4 localization and R-loop removal. REXO4 ablation-induced DNA damage stimulates interferon response and tumor immune infiltration to suppress squamous cell carcinoma (SCC) growth and metastasis. Importantly, targeting the exonuclease activity of REXO4 via the inhibitor we developed synergizes with PD-1 blockade to inhibit SCC progression by recruiting and activating CD8+ T cells. Thus, our study provides mechanistic insight into how m6A couples with exonuclease in R-loop clearance and genome maintenance, and uncovers a druggable epi-transcriptional machinery that blocks innate immune response and enables SCC immune evasion.

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RNA exonuclease REXO4 resolves m6A-marked R-loops and suppresses anti-tumor immunity [CUT&Tag-seq]

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RNA exonuclease REXO4 resolves m6A-marked R-loops and suppresses anti-tumor immunity

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