Fig. 5

Results of ‘RNA-induced DNA replication interference’ assays. a Effects of 27nt-RNAs using Pfu or Taq polymerases after end-point-PCR and agarose gel electrophoresis (top) or after qPCR (bottom). b Effects of 27nt-RNAs alone or in combination with PIWI1 on linear DNA amplification in a Klenow reaction were assayed via qPCR. c Hypothetical models on sequence-specific targeting through Argonaute/PIWI-RNA complexes (blue: IES, red: MDS, yellow: PIWI1, green: 27nt-RNA, orange: tethered transcript [c1 only]): c1 According to the ‚nascent transcript model, 27nt-RNA/PIWI1 complexes could target tethered IES-originating transcripts, which would be reminiscent of observations made in divergent eukaryotes, such as S. pombe, C. elegans and A. thaliana (reviewed in [38]). Alternatively, we assumed that 27nt-RNA/PIWI1 complexes could interact with dsDNA (c2) or via base-pairing with ssDNA, possibly when it occurs in a replication bubble (c3). d Mapping of mRNA reads purified 20 h PC on micronuclear model genes reveals that IES (red bars) are sharply omitted