Document Detail


Establishment of cells to monitor Microprocessor through fusion genes of microRNA and GFP.
MedLine Citation:
PMID:  18533106     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Microprocessor, the complex of Drosha and DGCR8, promotes the processing of primary microRNA to precursor microRNA, which is a crucial step for microRNA maturation. So far, no convenient assay systems have been developed for observing this step in vivo. Here we report the establishment of highly sensitive cellular systems where we can visually monitor the function of Microprocessor. During a series of screening of transfectants with fusion genes of the EGFP cDNA and primary microRNA genes, we have obtained certain cell lines where introduction of siRNA against DGCR8 or Drosha strikingly augments GFP signals. In contrast, these cells have not responded to Dicer siRNA; thus they have a unique character that GFP signals should be negatively and specifically correlated to the action of Microprocessor among biogenesis of microRNA. These cell lines can be useful tools for real-time analysis of Microprocessor action in vivo and identifying its novel modulators.
Authors:
Motomu Tsutsui; Hitoki Hasegawa; Koichi Adachi; Maiko Miyata; Peng Huang; Naoki Ishiguro; Michinari Hamaguchi; Takashi Iwamoto
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-06-03
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  372     ISSN:  1090-2104     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2008 Aug 
Date Detail:
Created Date:  2008-06-25     Completed Date:  2008-07-24     Revised Date:  2011-11-03    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  856-61     Citation Subset:  IM    
Affiliation:
Department of Orthopedic Surgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Sowa-ku, Nagoya, Aichi 466-8550, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
DEAD-box RNA Helicases / genetics
Endoribonucleases / genetics
Gene Fusion
Green Fluorescent Proteins / analysis,  genetics
Humans
Mice
MicroRNAs / genetics,  metabolism*
Molecular Sequence Data
NIH 3T3 Cells*
Proteins / genetics,  metabolism*
Ribonuclease III / genetics,  metabolism*
Transfection
Chemical
Reg. No./Substance:
0/DGCR8 protein, human; 0/MicroRNAs; 0/Proteins; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins; EC 3.1.-/Endoribonucleases; EC 3.1.26.3/DICER1 protein, human; EC 3.1.26.3/DROSHA protein, human; EC 3.1.26.3/Ribonuclease III; EC 3.6.1.-/DEAD-box RNA Helicases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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