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Small RNAseq analysis of C.elegans from Parental P0 to F3 generations under hypoxic stress in both WT and hrde-1 mutant [small RNA-seq]

ABSTRACT: The goal of small RNA-seq is to identify the differentially expressed small RNAs (microRNA, 22G RNA, piRNA, snoRNA) in the wild-type (WT) and hrde-1 mutant of C.elegans at the normoxia reared and hypoxia treatment conditions at P0 generation, and those small RNAs maintained dysregulation in F1, F2 and F3 generation.

ORGANISM(S): Caenorhabditis elegans

PROVIDER: GSE188270 | GEO | 2022/12/13

REPOSITORIES: GEO

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Transcriptome analysis in C.elegans from Parental P0 to F3 generations under hypoxic stress in both WT and hrde-1 mutant [RNA-seq]

Project description:The goal of RNA-seq is to identify the differentially expressed genes in the wild-type (WT) and hrde-1 mutant of C.elegans at the normoxia reared and hypoxia treatment conditions at P0 generation, and those maintained dysregulated genes in F1, F2 and F3 generation. Two biological replicates were assigned for each group and in total 32 groups were prepared for RNA-seq libraries with 500 ng mRNA as starting materials using NEXTflex Illumina qRNA-Seq Library Prep Kit (Bioo Scientific). We mapped about 20 - 30 million reads per sample to C.elegans (WS235) with RSEM pipline. Differential analysis were conducted using EdgeR. The significant genes were conducted by computing NB exact genewise tests with corrected p < 0.05.

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Germ granule association drives small RNA specificity for a nuclear Argonaute protein

Project description:RNA interference (RNAi) is a conserved gene silencing process that exists in diverse organisms to protect genome integrity and regulate gene expression. In C. elegans, the majority of RNAi pathway proteins localize to perinuclear, phase-separated germ granules, which are comprised of sub-domains referred to as P granules, Mutator foci, Z granules, and SIMR foci. However, the protein components and function of the newly discovered SIMR foci are unknown. Here we demonstrate that HRDE-2 localizes to SIMR foci and interacts with the germline nuclear RNAi Argonaute HRDE-1. Furthermore, HRDE-1 also localizes to SIMR foci, dependent on HRDE-2, but only in its small RNA unbound state. This germ granule localization is critical to promote the small RNA binding specificity of HRDE-1 and, in the absence of HRDE-2, HRDE-1 exclusively loads CSR-class 22G-RNAs rather than WAGO-class 22G-RNAs, resulting in H3K9me3 deposition on CSR-target genes. Thus, our study demonstrates that HRDE-2 is critical to ensure that the correct small RNAs are used to guide nuclear RNA silencing in the C. elegans germline.

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A nuclear Argonaute promotes germline immortality and epigenetic inheritance across generations

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Transcriptome and Small RNA analysis of C.elegans from Parental P0 to F3 generations under hypoxic stress in both WT and hrde-1 mutant

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