Construction and identification of different stem shRNA expression vectors / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 386-392, 2010.
Artigo
em Chinês
| WPRIM
| ID: wpr-336215
ABSTRACT
We constructed shRNA vectors with different stem length, and tested the silencing effectiveness in mouse cells and embryos. We designed interfering RNAs with stems of 21 bp, 27 bp, and 29 bp. The enhanced green fluorescent protein gene was used as target gene. The synthesized single strands were annealed and cloned into psiSTRIKE and the recombinant plasmids (EGFP-21 siRNA, EGFP-27 siRNA, and EGFP-29 siRNA) were transfected into the mouse embryonic fibroblast with lipofection. The mRNA expression level of the enhanced green fluorescent protein gene was checked by real-time quantitative PCR. The silencing effectiveness of the 29 bp shRNA vector was stronger than which of the 21 bp and 27 bp. The findings in this study are of interest for selecting the hairpins for mouse individuals.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Dados de Sequência Molecular
/
Sequência de Bases
/
Transfecção
/
Linhagem Celular
/
Caules de Planta
/
Inativação Gênica
/
Biologia Celular
/
RNA Interferente Pequeno
/
Proteínas de Fluorescência Verde
/
Embrião de Mamíferos
Tipo de estudo:
Estudo diagnóstico
/
Estudo prognóstico
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2010
Tipo de documento:
Artigo
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