Co-transfection and tandem transfection of HEK293A cells for overexpression and RNAi experiments.
Cell Biol Int
; 35(3): 187-92, 2011 Mar.
Article
em En
| MEDLINE
| ID: mdl-21087213
ABSTRACT
pIRES2-EGFP was employed and a non-target shRNA expressing plasmid was constructed to simulate overexpression and RNAi (RNA interference) experiments. Transfection of pIRES2-EGFP into HEK293A cells by cationic lipids VigoFect demonstrated that transfection efficiency increased in a dose-dependent manner with amount of DNA plasmid used, and optimal transfection time and cell density should be identified to reach a compromise of higher transfection efficiency and lower toxicity. Co-transfection experiments indicated that the two co-transfected plasmids were equivalently delivered into the same cells, and the co-transfection efficiency was rarely affected by cell density and proportion of the two plasmids. However, plasmid-receipted cells seemed indisposed to accept plasmid again during the second transfection, and very low co-transfection efficiency was observed in tandem transfection.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Transfecção
/
Interferência de RNA
Limite:
Humans
Idioma:
En
Revista:
Cell Biol Int
Ano de publicação:
2011
Tipo de documento:
Article