Comparison of approaches for efficient gene silencing induced by microRNA-based short hairpin RNA and indicator gene expression.
Mol Biol Rep
; 37(4): 1831-9, 2010 Apr.
Article
em En
| MEDLINE
| ID: mdl-19603286
ABSTRACT
MicroRNA-based short hairpin RNAs (shRNAs) are natural inducers of RNA interference and have been increasingly used in shRNA expression strategies. In the present study, we compared the efficiencies of exogenous green fluorescence protein (GFP) and endogenous glyceraldehyde-3-phosphate dehydrogenase (GAPDH) knockdown and red fluorescent protein (RFP) indicator expression mediated by three differently designed plasmids. RFP was introduced either at the 5' end, at the 3' end of the human mir155-based target gene (TG) (e.g., GFP or GAPDH) shRNA expression cassette (EC), or at the 3' end of the chimeric intron-containing TG shRNA EC. Comparisons with the control vector showed an obvious reduction of GFP or GAPDH expression with the various shRNA expression plasmids (P < 0.05). When RFP was located at the 5' end or at the 3' end of the TG shRNA EC, RFP expression was low; whereas when RFP was connected with the chimeric intron-containing TG shRNA EC, RFP expression was high. Taken together, this study demonstrated an efficient plasmid design for both TG silencing induced by microRNA-based shRNA and indicator gene expression in vitro.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Regulação da Expressão Gênica
/
Técnicas Genéticas
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Inativação Gênica
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MicroRNAs
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RNA Interferente Pequeno
Limite:
Humans
Idioma:
En
Revista:
Mol Biol Rep
Ano de publicação:
2010
Tipo de documento:
Article
País de afiliação:
China