Dataset Information


Phosphate-independent small RNA sequencing of WT and mir-249 mutant C. elegans

ABSTRACT: MicroRNAs (miRNA), discovered in C. elegans, are short non-coding RNAs that bind and regulate the expression of target mRNAs in animals and plants. C. elegans miRNAs bind to partially complementary sequences in the 3' UTR of the target mRNA, which results in translational repression through mRNA destabilization. The high-throughput sequencing of RNA cleavage fragments was performed to directly detect cleaved miRNA targets in C. elegans. Based on this analysis, miR-249 was identified as a potential miRNA that regulates a ZK637.6 pseudogene, paralogous to asna-1 (ZK637.5), by a cleavage mechanism with extensive, evolutionary conserved complementarity. Additionally, we validated miR-249-directed cleavage of the ZK637.6 by a gene-specific 5’ rapid amplification of cDNA ends and observed notable difference in expression of ZK637.6 in wild-type versus mir-249 mutant C. elegans. Furthermore, phosphate-independent small-RNA sequencing analysis revealed that miR-249 target genes, including ZK637.6 pseudogene, showed 22G-RNA productions dependent on miR-249 targeting. These findings may lead to a better understanding of the biological roles of miRNAs for pseudogenes in C. elegans. Total small RNAs from wild-type and mir-249 mutant in adult stage worms were subjected to small RNA sequencing using an Illumina platform with Tobacco Acid Pyrophosphatase (TAP) treatment, allowing detection of secondary siRNAs carrying 5’-tri-phosphate.

ORGANISM(S): Caenorhabditis elegans  

SUBMITTER: Jin W Nam   June H Park  Chanseok Shin 

PROVIDER: E-GEOD-41900 | ArrayExpress | 2013-08-08



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Degradome sequencing reveals an endogenous microRNA target in C. elegans.

Park June Hyun JH   Ahn Soungyub S   Kim Soyoung S   Lee Junho J   Nam Jin-Wu JW   Shin Chanseok C  

FEBS letters 20130226 7

Caenorhabditis elegans microRNAs (miRNAs) bind to partially complementary sequences in the 3' untranslated region of target mRNAs, resulting in translational repression through mRNA destabilization. High-throughput sequencing of RNA cleavage fragments was performed to directly detect miRNA-directed cleavage targets in adult stage C. elegans. From this analysis, we found that miR-249 directed the cleavage of the ZK637.6 transcript with extensive and evolutionarily conserved complementarity in nem  ...[more]

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