AI Article Synopsis
- Recent research indicates that transcription occurs at DNA double-strand breaks (DSBs) and that small RNAs, called damage-induced small RNAs (diRNAs), play a crucial role in DNA repair.
- The study employed the homing endonuclease I-PpoI to explore diRNA production in human and mouse cells, revealing that significant levels of diRNAs are generated specifically from the repetitive 28S rRNA locus, while unique and intergenic loci produce little to none.
- The findings challenge the necessity of certain RNA processing proteins for diRNA production, showing that Drosha and DGCR8 are not essential, and highlight the presence of two distinct diRNA subtypes that are incorporated into Argonaute protein complexes.
Article Abstract
Recent studies suggest that transcription takes place at DNA double-strand breaks (DSBs), that transcripts at DSBs are processed by Drosha and Dicer into damage-induced small RNAs (diRNAs), and that diRNAs are required for DNA repair. However, diRNAs have been mostly detected in reporter constructs or repetitive sequences, and their existence at endogenous loci has been questioned by recent reports. Using the homing endonuclease I-PpoI, we have investigated diRNA production in genetically unperturbed human and mouse cells. I-PpoI is an ideal tool to clarify the requirements for diRNA production because it induces DSBs in different types of loci: the repetitive 28S locus, unique genes and intergenic loci. We show by extensive sequencing that the rDNA locus produces substantial levels of diRNAs, whereas unique genic and intergenic loci do not. Further characterization of diRNAs emerging from the 28S locus reveals the existence of two diRNA subtypes. Surprisingly, Drosha and its partner DGCR8 are dispensable for diRNA production and only one diRNAs subtype depends on Dicer processing. Furthermore, we provide evidence that diRNAs are incorporated into Argonaute. Our findings provide direct evidence for diRNA production at endogenous loci in mammalian cells and give insights into RNA processing at DSBs.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6294500 | PMC |
| http://dx.doi.org/10.1093/nar/gky1107 | DOI Listing |
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