Unknown,Transcriptomics,Genomics,Proteomics

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Characterization of the piRNA Complex from Rat Testes


ABSTRACT: This data series contains the sequences and read counts of Piwi-interacting RNAs and other small RNAs from mouse and rat testes extracts. In a search for different classes of small RNAs that might be involved in transcriptional gene silencing, we encountered a novel class of small RNAs within mammalian testes. This study reports the identification of small RNAs and their relative abundance levels to each other based on the deep coverage of the cDNA library. Keywords: High-throughput pyrosequencing by the 454 Life Sciences Genome Sequencer 20™ System We constructed a cDNA library from small RNAs purified from Q column fractions of testes extracts. Briefly, gel-purified small RNAs from the column fractions were ligated with adaptor oligos on both 3' and 5' ends of the small RNAs. Reverse-transcription and PCR amplification was then performed on the ligated small RNAs to yield a cDNA library. This cDNA library was finally converted by PCR into a single-stranded DNA library compatible for coupling to beads on the 454 Life Sciences Genome Sequencer 20™ System. Reads and sequences of the small RNAs were processed from linker sequences, examined for matches to the mouse or rat genomes, and tallied.

ORGANISM(S): Mus musculus

SUBMITTER: Nelson Lau 

PROVIDER: E-GEOD-5026 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Characterization of the piRNA complex from rat testes.

Lau Nelson C NC   Seto Anita G AG   Kim Jinkuk J   Kuramochi-Miyagawa Satomi S   Nakano Toru T   Bartel David P DP   Kingston Robert E RE  

Science (New York, N.Y.) 20060615 5785


Small noncoding RNAs regulate processes essential for cell growth and development, including mRNA degradation, translational repression, and transcriptional gene silencing (TGS). During a search for candidate mammalian factors for TGS, we purified a complex that contains small RNAs and Riwi, the rat homolog to human Piwi. The RNAs, frequently 29 to 30 nucleotides in length, are called Piwi-interacting RNAs (piRNAs), 94% of which map to 100 defined (< or = 101 kb) genomic regions. Within these re  ...[more]

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