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Production of artificial piRNAs in flies and mice

Muerdter, Felix and Olovnikov, Ivan and Molaro, Antoine and Rozhkov, Nikolay V. and Czech, Benjamin and Gordon, Azzaf and Hannon, Gregory J. and Aravin, Alexei A. (2012) Production of artificial piRNAs in flies and mice. RNA, 18 (1). pp. 42-52. ISSN 1355-8382. PMCID PMC3261743.

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In animals a discrete class of small RNAs, the piwi-interacting RNAs (piRNAs), guard germ cell genomes against the activity of mobile genetic elements. piRNAs are generated, via an unknown mechanism, from apparently single-stranded precursors that arise from discrete genomic loci, termed piRNA clusters. Presently, little is known about the signals that distinguish a locus as a source of piRNAs. It is also unknown how individual piRNAs are selected from long precursor transcripts. To address these questions, we inserted new artificial sequence information into piRNA clusters and introduced these marked clusters as transgenes into heterologous genomic positions in mice and flies. Profiling of piRNA from transgenic animals demonstrated that artificial sequences were incorporated into the piRNA repertoire. Transgenic piRNA clusters are functional in non-native genomic contexts in both mice and flies, indicating that the signals that define piRNA generative loci must lie within the clusters themselves rather than being implicit in their genomic position. Comparison of transgenic animals that carry insertions of the same artificial sequence into different ectopic piRNA-generating loci showed that both local and long-range sequence environments inform the generation of individual piRNAs from precursor transcripts.

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Additional Information:© 2012 RNA Society. Published by Cold Spring Harbor Laboratory Press. Received August 8, 2011. Accepted September 26, 2011. Published in Advance November 17, 2011. We thank members of the Hannon and Aravin labs for helpful discussion and comments on the manuscript. We thank members of the McCombie lab (CSHL) and Igor Antoshechkin (Caltech) for help with RNA sequencing. We thank Andres Canela (CSHL) for technical assistance and Simon Knott (CSHL) and Alex Zahn (Caltech) for help with statistical analysis. Sang Yong Kim (CSHL) created the transgenic mice used in this study. F.M. was supported by the Volkswagen Foundation and B.C. by the Boehringer Ingelheim Fonds. This work was supported by grants from the National Institutes of Health (DP2 OD007371A and R00HD057233 to A.A.A.; 5R01GM062534 to G.J.H.), by the Ellison Medical Foundation (A.A.A.), and by a kind gift from Kathryn W. Davis (G.J.H.).
Funding AgencyGrant Number
Volkswagen FoundationUNSPECIFIED
Boehringer Ingelheim FondsUNSPECIFIED
NIHDP2 OD007371A
NIHR00 HD057233
Ellison Medical FoundationUNSPECIFIED
Subject Keywords:piwi; noncoding RNA; piRNA
Issue or Number:1
PubMed Central ID:PMC3261743
Record Number:CaltechAUTHORS:20120126-110548127
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Official Citation:Production of artificial piRNAs in flies and mice Felix Muerdter, Ivan Olovnikov, Antoine Molaro, Nikolay V. Rozhkov, Benjamin Czech, Assaf Gordon, Gregory J. Hannon, and Alexei A. Aravin RNA January 2012 18: 42-52; Published in Advance November 17, 2011, doi:10.1261/rna.029769.111
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:28982
Deposited By: Ruth Sustaita
Deposited On:26 Jan 2012 19:43
Last Modified:03 Oct 2019 03:37

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