[Construction of VEGF gene-targeted small interfering RNA mediated by CMV promoter and its silencing effect].
Zhejiang Da Xue Xue Bao Yi Xue Ban
; 39(2): 181-6, 2010 03.
Article
em Zh
| MEDLINE
| ID: mdl-20387247
ABSTRACT
OBJECTIVE:
To construct VEGF gene-targeted small interfering RNA (siRNA) and its expression vector driven by CMV promoter and to investigate its interference effect.METHODS:
The VEGF gene-targeted hairpin siRNA was designed, two complementary oligonucleotide strands were synthesized. After annealing, two-strand oligonucleotide was inserted into pDC311-SV40-RC vector, which was then identified by PCR and sequenced. Then human U-2 OS cell line was transfected with the vector using lipofectamine method. Finally, ELISA was performed to evaluate the expression of VEGF protein.RESULTS:
PCR-identification of positive clone and sequencing confirmed the vector containing the target siRNA. ELISA showed that compared with the control group, the expression levels of VEGF protein in transfected U-2 OS cells were decreased significantly (P<0.05).CONCLUSION:
VEGF gene-targeted siRNA and its vector mediated by CMV promoter were successfully constructed, which can reduce the VEGF protein expression after transfecting.
Buscar no Google
Base de dados:
MEDLINE
Assunto principal:
Regiões Promotoras Genéticas
/
Inativação Gênica
/
RNA Interferente Pequeno
/
Fator A de Crescimento do Endotélio Vascular
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
Zh
Ano de publicação:
2010
Tipo de documento:
Article