Document Detail


RNPomics: defining the ncRNA transcriptome by cDNA library generation from ribonucleo-protein particles.
MedLine Citation:
PMID:  20150415     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Up to 450,000 non-coding RNAs (ncRNAs) have been predicted to be transcribed from the human genome. However, it still has to be elucidated which of these transcripts represent functional ncRNAs. Since all functional ncRNAs in Eukarya form ribonucleo-protein particles (RNPs), we generated specialized cDNA libraries from size-fractionated RNPs and validated the presence of selected ncRNAs within RNPs by glycerol gradient centrifugation. As a proof of concept, we applied the RNP method to human Hela cells or total mouse brain, and subjected cDNA libraries, generated from the two model systems, to deep-sequencing. Bioinformatical analysis of cDNA sequences revealed several hundred ncRNP candidates. Thereby, ncRNAs candidates were mainly located in intergenic as well as intronic regions of the genome, with a significant overrepresentation of intron-derived ncRNA sequences. Additionally, a number of ncRNAs mapped to repetitive sequences. Thus, our RNP approach provides an efficient way to identify new functional small ncRNA candidates, involved in RNP formation.
Authors:
Mathieu Rederstorff; Stephan H Bernhart; Andrea Tanzer; Marek Zywicki; Katrin Perfler; Melanie Lukasser; Ivo L Hofacker; Alexander Hüttenhofer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-02-11
Journal Detail:
Title:  Nucleic acids research     Volume:  38     ISSN:  1362-4962     ISO Abbreviation:  Nucleic Acids Res.     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-06-03     Completed Date:  2010-06-22     Revised Date:  2010-09-28    
Medline Journal Info:
Nlm Unique ID:  0411011     Medline TA:  Nucleic Acids Res     Country:  England    
Other Details:
Languages:  eng     Pagination:  e113     Citation Subset:  IM    
Affiliation:
Division of Genomics and RNomics, Innsbruck Biocentre, Innsbruck Medical University, Innsbruck and Institute of Theoretical Chemistry, University of Vienna, Vienna, Austria.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Brain Chemistry
Conserved Sequence
Exons
Gene Expression Profiling
Gene Library*
Hela Cells
Humans
Introns
Mice
RNA, Untranslated / classification,  isolation & purification,  metabolism*
Repetitive Sequences, Nucleic Acid
Ribonucleoproteins / chemistry*,  isolation & purification
Chemical
Reg. No./Substance:
0/RNA, Untranslated; 0/Ribonucleoproteins
Comments/Corrections

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