Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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A variety of Dicer substrates in human and C. elegans (HEK small RNA)


ABSTRACT: The endoribonuclease Dicer is known for its central role in the biogenesis of eukaryotic small RNAs/microRNAs. Despite its importance, Dicer target transcripts have not been directly mapped. Here, we apply biochemical methods to human cells and C. elegans and identify thousands of Dicer binding sites. We find known and hundreds of novel miRNAs with high sensitivity and specificity. We also report structural RNAs, promoter RNAs, and mitochondrial transcripts as Dicer targets. Interestingly, most Dicer binding sites reside on mRNAs/lncRNAs and are not significantly processed into small RNAs. These passive sites typically harbor small, Dicer-bound hairpins within intact transcripts and generally stabilize target expression. We show that passive sites can sequester Dicer and reduce microRNA expression. mRNAs with passive sites were in human and worm significantly associated with processing-body/granule function. Together, we provide the first transcriptome-wide map of Dicer targets and suggest conserved binding modes and functions outside the miRNA pathway. small RNAs were extracted from total RNA. Standard small RNA sequencing was performed to sequence 5' monophosphate bearing small RNAs. This was performed for siRNA knockdowns of DICER1 or DROSHA, with two independent siRNAs each, and for control cells. (5 samples in total)

ORGANISM(S): Homo sapiens

SUBMITTER: Marvin Jens 

PROVIDER: E-GEOD-63458 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

A variety of dicer substrates in human and C. elegans.

Rybak-Wolf Agnieszka A   Jens Marvin M   Murakawa Yasuhiro Y   Herzog Margareta M   Landthaler Markus M   Rajewsky Nikolaus N  

Cell 20141101 5


The endoribonuclease Dicer is known for its central role in the biogenesis of eukaryotic small RNAs/microRNAs. Despite its importance, Dicer target transcripts have not been directly mapped. Here, we apply biochemical methods to human cells and C. elegans and identify thousands of Dicer-binding sites. We find known and hundreds of additional miRNAs with high sensitivity and specificity. We also report structural RNAs, promoter RNAs, and mitochondrial transcripts as Dicer targets. Interestingly,  ...[more]

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