Comment[ArrayExpressAccession] E-MTAB-1557 MAGE-TAB Version 1.1 Investigation Title Multiple epigenetic mechanisms and the piRNA pathway enforce LINE1 silencing during adult spermatogenesis Comment[Submitted Name] Multiple epigenetic mechanisms and the piRNA pathway enforce LINE1 silencing during adult spermatogenesis Experiment Description The Piwi protein Mili and the Piwi-interacting RNA (piRNA) pathway are required to establish L1 DNA methylation during epigenetic reprogramming that accompanies male germ cell development. Although Mili is expressed throughout most of adult spermatogenesis, its function therein remains unknown. We find that Mili displays a hitherto unappreciated dynamic expression profile during spermatogenesis, expressed in the mitotic spermatogonia but absent at the early meiotic stages only to reappear in late zygotene spermatocytes. Here we show using conditional mutagenesis that Mili's endonuclease activity post-transcriptionally cleaves L1 transcripts commencing upon its reappearance in late zygotene spermatocytes even in the presence of normal L1 DNA methylation Experimental Design individual_genetic_characteristics_design genetic_modification_design in_vivo_design replicate_design microRNA profiling by high throughput sequencing Comment[AEExperimentType] RNA-seq of non coding RNA Comment[AEExperimentDisplayName] RNA-seq of non coding RNA of mouse Mili_Ctl and Mili_C-DAH testes to investigate epigenetic mechanisms and the piRNA pathway enforce LINE1 silencing during adult spermatogenesis Experimental Factor Name immunoprecipitate genotype Experimental Factor Type immunoprecipitate genotype Quality Control Type biological_replicate Quality Control Term Source REF EFO Public Release Date 2013-07-03 Person Last Name Saini Person First Name Harpreet Person Mid Initials K Person Email hsaini@ebi.ac.uk Person Phone +44-1223-492676 Person Address EMBL-EBI, Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SD, United Kingdom Person Affiliation EMBL-EBI Person Roles submitter PubMed ID 23706823 Publication Author List Monica Di Giacomo, Stefano Comazzetto, Harpreet Saini, Serena De Fazio, Claudia Carrieri, Marcos Morgan, Lina Vasiliauskaite, Vladimir Benes, Anton J. Enright, Donal O'Carroll Publication Title Multiple epigenetic mechanisms and the piRNA pathway enforce LINE1 silencing during adult spermatogenesis Publication Status submitted Protocol Name P-MTAB-22753 P-MTAB-31669 P-MTAB-31670 Protocol Type nucleic acid sequencing protocol high throughput sequence alignment protocol nucleic acid library construction protocol Protocol Description Standard Illumina clustering and sequencing protocol for Illumina GAIIx sequencer were used. The reads were strippped of adapters and linkers, and low quality regions were masked. Only reads between length 24 and 30 were kept and aligned to mm9 genome using Bowtie. Small RNA libraries were generated with the protocol described in Hafner M. et al. (2008) but using adaptors suitable for sequencing on the Ilumina platform. Protocol Hardware Illumina Genome Analyzer IIx Protocol Term Source REF ArrayExpress EFO EFO Term Source Name EFO ArrayExpress Term Source File http://www.ebi.ac.uk/efo http://www.ebi.ac.uk/ArrayExpress Comment[SecondaryAccession] ERP002365 Comment[SequenceDataURI] http://www.ebi.ac.uk/ena/data/view/ERR245087-ERR245090 SDRF File E-MTAB-1557.sdrf.txt Publication DOI 10.1016/j.molcel.2013.04.026