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.
Artigo
em Chinês
| WPRIM
| ID: wpr-340779
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>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
RNA Mensageiro
/
Transfecção
/
Regulação para Baixo
/
Linhagem Celular
/
Biologia Celular
/
Miócitos Cardíacos
/
MicroRNAs
/
RNA Interferente Pequeno
/
Proteínas de Fluorescência Verde
Tipo de estudo:
Estudo prognóstico
Limite:
Animais
Idioma:
Chinês
Revista:
Journal of Southern Medical University
Ano de publicação:
2008
Tipo de documento:
Artigo
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