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Series GSE80771 Query DataSets for GSE80771
Status Public on Sep 01, 2016
Title A sliding-bulge structure at the Dicer processing site of pre-miRNA regulates alternative Dicer processing to generate 5’-isomiRs (pre-miR-203)
Organism Homo sapiens
Experiment type Non-coding RNA profiling by high throughput sequencing
Summary 5’-isomiRs expand the repertoire of miRNA targets. However, how they are generated is not well understood. Here, we showed that, for some miRNAs in mammalian cells, 5’-isomiRs are generated by alternative Dicer processing by using miRNA offset RNAs (moRs) to determine Drosha cleavage sites in cells. In addition, we showed that in miR-203, alternative Dicer processing is regulated by a conserved sliding-bulge structure at the Dicer processing site, which allows the pre-miRNA molecule to fold into two different structures that are processed differently by Dicer. So far no RNA motif that slides to change conformation and alter a protein–RNA interaction has been reported. Thus, our study revealed a novel RNA motif that regulates 5’-isomiR generation in some miRNAs. It might also contribute to regulating protein–RNA interactions in other biological processes, since it takes only one point mutation to generate the sliding bulge, and there are a large number of different RNAs in the cell.
 
Overall design Mutants as well as the wild type of pri-miR-203 were designed. They were constructed into pLL3.7 and transfected into 293FT cells. The small RNAs were then profiled with deep sequencing. Three independent profiling exprements (replicates) were included in the dataset. RNA sample from each mutant of each replicate was barcoded, multicomplexed and sequenced in a single run of illumina Miseq sequencing.
 
Contributor(s) Wu H, Ma H
Citation(s) 27656682
Submission date Apr 28, 2016
Last update date May 15, 2019
Contact name Haoquan wu
E-mail(s) haoquan.wu@ttuhsc.edu
Phone 915-2154243
Organization name Texas Tech Health Sciences Center El Paso
Department Biomedical Scienes
Lab COE-ID
Street address 5001 El Paso Dr
City El Paso
State/province Texas
ZIP/Postal code 79905
Country USA
 
Platforms (1)
GPL15520 Illumina MiSeq (Homo sapiens)
Samples (2)
GSM2136841 pre-miR-203_old mutants
GSM2136842 pre-miR-203_new mutants
Relations
BioProject PRJNA319961
SRA SRP074124

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Supplementary file Size Download File type/resource
GSE80771_RAW.tar 60.0 Kb (http)(custom) TAR (of XLSX)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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