A genome-wide RNAi screen identifies factors required for distinct stages of C-elegans piRNA biogenesis

Goh, W. S. S., Seah, J. W. E., Harrison, E. J., Chen, C., Hammell, C. M., Hannon, G. J. (April 2014) A genome-wide RNAi screen identifies factors required for distinct stages of C-elegans piRNA biogenesis. Genes & Development, 28 (7). pp. 797-807. ISSN 0890-9369

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URL: http://www.ncbi.nlm.nih.gov/pubmed/24696458
DOI: 10.1101/gad.235622.113

Abstract

In animals, piRNAs and their associated Piwi proteins guard germ cell genomes against mobile genetic elements via an RNAi-like mechanism. In Caenorhabditis elegans, 21U-RNAs comprise the piRNA class, and these collaborate with 22G RNAs via unclear mechanisms to discriminate self from nonself and selectively and heritably silence the latter. Recent work indicates that 21U-RNAs are post-transcriptional processing products of individual transcription units that produce similar to 26-nucleotide capped precursors. However, nothing is known of how the expression of precursors is controlled or how primary transcripts give rise to mature small RNAs. We conducted a genome-wide RNAi screen to identify components of the 21U biogenesis machinery. Screening by direct, quantitative PCR (qPCR)-based measurements of mature 21U-RNA levels, we identified 22 genes important for 21U-RNA production, termed TOFUs (Twenty-One-u Fouled Ups). We also identified seven genes that normally repress 21U production. By measuring mature 21U-RNA and precursor levels for the seven strongest hits from the screen, we assigned factors to discrete stages of 21U-RNA production. Our work identifies for the first time factors separately required for the transcription of 21U precursors and the processing of these precursors into mature 21U-RNAs, thereby providing a resource for studying the biogenesis of this important small RNA class.

Item Type: Paper
Subjects: organism description > animal > C elegans
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNAi
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > piRNA
CSHL Authors:
Communities: CSHL Cancer Center Program > Cancer Genetics
CSHL Cancer Center Shared Resources > Bioinformatics Service
CSHL labs > Hammell C. lab
CSHL labs > Hannon lab
Watson School > Publications
CSHL Cancer Center Shared Resources > DNA Sequencing Service
Depositing User: Matt Covey
Date: April 2014
Date Deposited: 15 May 2014 19:26
Last Modified: 04 Nov 2015 17:14
PMCID: PMC4015493
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30136

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