Construction and identification of a novel adeno-integrase hybrid system for hemophilia B / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 1229-1234, 2010.
Article
en Zh
| WPRIM
| ID: wpr-332387
Biblioteca responsable:
WPRO
ABSTRACT
This study was aimed to construct an adenovirus hybrid system with high transduction efficiency and site-specific integration. By a series of DNA manipulation, a hybrid system of two adenovirus vectors was constructed. One vector contains loxP-flanked transgene expression cassette, in which there are hFIX and DsRed coding sequences and attB for phiC31 recolonization. The other vector carries Cre and phiC31 gene. Vectors only expressing Cre or phiC31 were used as controls. 293A cells were constructed and transfected with the adenoviral vectors by Lipofectamine 2000, and the expression of target genes was identified by fluorescence microscopy and RT-PCR. The results showed that after being identified by PCR, restriction analysis and sequencing, an adeno-integrase hybrid system was successfully constructed. The system expressed RFP, GFP, hFIX, Cre and phiC31 in 293A cells in vitro. It is concluded that the adeno-integrase hybrid system is successfully constructed, which lays a good foundation for further investigation of its therapeutic application.
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Terapéutica
/
Transfección
/
Terapia Genética
/
Expresión Génica
/
Línea Celular
/
Adenoviridae
/
Hemofilia B
/
Técnicas de Transferencia de Gen
/
Integrasas
/
Vectores Genéticos
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Límite:
Humans
Idioma:
Zh
Revista:
Journal of Experimental Hematology
Año:
2010
Tipo del documento:
Article