RT Journal Article
SR Electronic
T1 siRNAs targeting a chromatin-associated RNA induce its transcriptional silencing in human cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.05.11.491477
DO 10.1101/2022.05.11.491477
A1 Julien Ouvrard
A1 Lisa Muniz
A1 Estelle Nicolas
A1 Didier Trouche
YR 2022
UL http://biorxiv.org/content/early/2022/05/11/2022.05.11.491477.abstract
AB Transcriptional gene silencing by small interfering RNAs (siRNAs) has been widely described in various species, such as plants or fission yeasts. In mammals, its extent remained somewhat debated. Previous studies showed that siRNAs targeting gene promoters can induce the silencing of the targeted promoter, although the involvement of off-target mechanisms was also suggested. Here, by nascent RNA capture and RNA Pol II ChIP, we show that siRNAs targeting non coding RNAs that are mainly localized in chromatin induce their transcriptional silencing. Moreover, by deleting the targeted sequence on the two alleles of one of these siRNAs by genome edition, we further found that this silencing is due to base pairing of the siRNA to the target. By using cells in which the deletion of the targeted sequence is heterozygous, we show by allele-specific RT-qPCR that only the wild type allele, but not the deleted allele, is silenced by the siRNA, indicating that transcriptional silencing occurs only in cis. Finally, we demonstrate that both Ago1 and Ago2 are involved in VINK transcriptional silencing. Altogether, our data demonstrate that siRNAs targeting chromatin-associated RNAs at distance from their promoter induce their gene silencing. Our results thus extend the possible repertoire of endogenous or exogenous interfering RNAs.Competing Interest StatementThe authors have declared no competing interest.