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The mechanism that elicits the formation of these non-canonical RNA:DNA structures is yet unclear although they were proposed to arise consequently to RNA Polymerase II or III loading at the break site followed by DSB-induced de novo transcription. Here, we found that none of these RNA polymerases are recruited at DSBs. Rather, strand-specific R-loop mapping revealed that RNA:DNA hybrids are mainly generated from the hybridization of pre-existing pre-mRNA to the 3’ overhang left by DNA end resection. We further identified the H3K4me3 reader Spindlin 1 to be essential for RNA:DNA hybrids accumulation at DSBs, through its role in mediating transcriptional repression in cis to DSBs. Altogether, we provide evidence that RNA:DNA hybrids accumulate at DSBs occurring in transcribing loci as a result of DSB-induced transcriptional shut-down."],"repository":["ENA"],"additional_accession":[]},"is_claimable":false,"name":"Transcriptional repression accounts for RNA:DNA hybrids accumulation at DNA Double Strand breaks","description":"Transcriptional repression accounts for RNA:DNA hybrids accumulation at DNA Double Strand breaks","dates":{"last_updated":"2025-06-29","first_public":"2025-06-29"},"accession":"PRJEB64562","cross_references":{}}