title: Production of artificial piRNAs in flies and mice.
abstract: 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.
authors.affiliation: Howard Hughes Medical Institute, Watson School of Biological Sciences, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.
authors.forename: Felix
authors.initials: F
authors.lastname: Muerdter
authors.forename: Ivan
authors.initials: I
authors.lastname: Olovnikov
authors.forename: Antoine
authors.initials: A
authors.lastname: Molaro
authors.forename: Nikolay V
authors.initials: NV
authors.lastname: Rozhkov
authors.forename: Benjamin
authors.initials: B
authors.lastname: Czech
authors.forename: Assaf
authors.initials: A
authors.lastname: Gordon
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Hannon
authors.forename: Alexei A
authors.initials: AA
authors.lastname: Aravin
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D034741:RNA, Small Interfering / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
chemical_list: D034741:RNA, Small Interfering
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