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SRX19477716: GSM7059825: Dmoj.ovary.1; Drosophila mojavensis; RNA-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 15.9M spots, 1.6G bases, 469.3Mb downloads

External Id: GSM7059825_r1
Submitted by: Hannon, Cancer Research UK - Cambridge Institute, University of Cambridge
Study: Unistrand piRNA clusters are an evolutionarily conserved mechanism to suppress endogenous retroviruses across the Drosophila genus [RNA-Seq]
show Abstracthide Abstract
The PIWI-interacting RNA (piRNA) pathway plays a crucial role in preventing endogenous genomic parasites, transposable elements (TEs), from damaging the genetic material of animal gonadal cells. Specific regions in the genome, called piRNA clusters, define each species' piRNA repertoire and therefore its capacity to recognize and silence transposons. In the somatic cells of the Drosophila melanogaster ovary, the flamenco (flam) unistrand cluster is the main source of piRNAs and primarily regulates Gypsy family TEs that are able to form virus-like particles and infect neighbouring germ cells. Disruption of the flam locus or failure to process flam precursor transcripts into piRNAs results in sterility, yet it remains unknown whether this silencing mechanism is employed widely across Drosophilidae. Here, using both synteny-based analyses and de novo TE annotation, we identify candidate loci sharing both their organisation and TE targeting repertoire with flam in widely divergent Drosophila species groups. Small RNA-sequencing validated these loci as bona-fide unistrand piRNA clusters and revealed their predominant expression in somatic cells of the ovary, likely to counter TE mobilisation in this tissue. This study provides compelling evidence of co-evolution between virus-like Gypsy family transposons and a host defence mechanism in form of soma-expressed, unistrand piRNA clusters. Overall design: Total RNA-seq from dissected ovaries from 10 Drosophila species, 1-8 replicates per species.
Sample: Dmoj.ovary.1
SAMN33419379 • SRS16869085 • All experiments • All runs
Library:
Name: GSM7059825
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Ovaries from 10-20 flies were dissected in ice-cold PBS and total RNA was extracted using TRIzol. Briefly, ovaries were lysed in TRIzol (Thermo Fisher Scientific), and RNA was phase separated using chloroform (Sigma-Aldrich). Total RNA in the aqueous fraction was precipitated using 100% isopropanol, washed with 70% ethanol, and dissolved in nuclease free water (Invitrogen). Ribosomal RNA was depleted using RiboPOOL (siTOOLs, Biotech) following the manufacturer's protocol. The indicated RiboZero libraries were prepared as previously described (Kneuss et al., 2019, PMID: 31416967). RNA-seq libraries were produced using NEBNext Ultra Directional Library Prep Kit for Illumina, following the manufacturer's instructions for rRNA depleted RNA.
Runs: 1 run, 15.9M spots, 1.6G bases, 469.3Mb
Run# of Spots# of BasesSizePublished
SRR2359263715,855,9531.6G469.3Mb2023-09-26

ID:
26756572

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