Plasmid-mediated miRNA-1-2 specifically inhibits Hand2 protein expression in H9C2 cells / 南方医科大学学报
Journal of Southern Medical University
; (12): 1559-1567, 2008.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-340779
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct an eukaryotic expression vector for miRNA-1-2 that can be expressed in rat H9C2 cardiomyocytes.</p><p><b>METHODS</b>The precursor miRNA (pre-miRNA) DNA template for miRNA-1-2 was designed and generated by PCR amplification. The DNA template was inserted into the hairpin RNA expression vector pSilence-4.1-neo and identified by DNA sequencing analysis. The recombinant plasmid DNA was then transfected into H9C2 cells via Lipofectamine, and the green fluorescence protein expression vector pEGFP-N3 served as the transfection marker. Twenty-four hours after transfection, the total cellular RNA was extracted using TRIzol reagent, and thermoscript reverse transcriptase (RT)-PCR was performed to determine miRNA-1-2 precursor expression.</p><p><b>RESULTS AND CONCLUSION</b>DNA sequencing indicated that the miR-1-2 expression plasmid was correctly constructed. The precursor miRNA-1-2 was successfully expressed in the H9C2 cells, and the expression of Hand2 protein could be efficiently inhibited by miRNA-1.</p>
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
RNA, Messenger
/
Transfection
/
Down-Regulation
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Cell Line
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Cell Biology
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Myocytes, Cardiac
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MicroRNAs
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RNA, Small Interfering
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Green Fluorescent Proteins
Type of study:
Prognostic study
Limits:
Animals
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2008
Document type:
Article