Gene expression of AAV-ITR ssDNA mini vector in skeletal muscle of mice.
- Author:
Dongqin ZHU
;
Yun ZHANG
;
Xiaomei LIU
;
Chun ZHANG
- Publication Type:Journal Article
- MeSH:
Animals;
DNA, Single-Stranded;
genetics;
Dependovirus;
genetics;
Gene Expression;
Genetic Therapy;
Genetic Vectors;
Mice;
Mice, Inbred ICR;
Muscle, Skeletal;
metabolism;
Real-Time Polymerase Chain Reaction
- From:
Chinese Journal of Biotechnology
2014;30(11):1720-1732
- CountryChina
- Language:Chinese
-
Abstract:
AAV-ITR single strand DNA mini vector (AAV-ITR ssDNA mini vector) is a novel gene expression vector based on AAV-ITR. We have shown efficient gene expression of AAV-ITR ssDNA mini vector in HEK 293T. Here, we studied the efficacy of gene expression of AAV-ITR ssDNA mini vector in vivo. We injected the skeletal muscle of ICR mice separately with equal molars of AAV-ITR ssDNA mini vector, ITR mutated AAV-ITR single strand DNA mini vector (AAV-ITRmm ssDNA mutant vector), AAV-ITR dsDNA and pUC57-minivector-GFP, combined with TurboFect. Florescence microscope analysis of skeletal muscle section shows that AAV-ITR ssDNA mini vector had higher expression efficiency and longer expression period. We extracted DNA from the muscle three months after injection and quantified three vectors by Real-time PCR. RT-PCR analysis shows that there were highest copy numbers of AAV-ITR ssDNA mini vector existing in muscle. Stable existing of AAV- TR ssDNA mini vector in muscle could be the molecular basis of long term gene expression of the vector. The results suggest that AAV-ITR ssDNA mini vector might be a promising vector for gene therapy.
