Figure S1. Two Independent CLASH Datasets, Related to Figure 1(A) Schematic of the CRISPR-engineered gfp::tev::flag::
prg-1 (gtf::
prg-1) gene (top), and fluorescence micrographs (bottom) of GFP::TEV::FLAG::PRG-1 (green)expression in the germline. PGL-1::mRFP (red) is a constitutive P granule marker.(B) Schematic of the gfp::
cdk-1 transgene (top), and fluorescence micrographs (bottom) of GFP::CDK-1 (green) expression in oocyte nuclei of wild-type (WT),
prg1(tm872), and gtf::
prg-1 worms.
prg-1(
tm872) is a loss-of-function allele that activates GFP::CDK-1 expression in (arrowheads). The GTF::PRG-1 fusion isfunctional.(C and D) The graphs present the number of reads mapped to each identified piRNA (C) and mRNA (D) in one experiment relative to the other one. r, Pearson'scorrelation coefficient.(E and F) Venn diagram summarizing the overlap in the number of piRNAs (E) and mRNA (F) in two replicates.(G and H) Distribution of all piRNA interactions among various types of RNAs in two replicates. mRNAs are the main piRNA targets and represent more than 70%.(I) Correlation between the abundance of CLASH-recovered piRNAs (x axis) and overall piRNA abundance (y axis).(J) Histogram showing the number of nucleotides added to the 50 (negative) or 30 (positive) ends of CLASH-recovered target regions to create ''ideal'' piRNA/targetRNA pairs. ''0'' indicates that no nucleotide was added to the target sequence. Hybrids ligated to the 30 ends of piRNAs (blue), and hybrids ligated to the 50 ends ofpiRNAs (red).(K) Distribution of chimeric
rrn-2.1 and
rrn-3.1 reads (green) identified by CLASH and genomic locus on the top (blue).(L) Putative interactions between piRNA and tRNA. 21ur-8377 reproducibly bind to the same region of tRNA-Glu(CUC), marked blue on the tRNA structure (chrIII.ZK783.
t1).