Recombinant adeno-associated virus type 8 mediated dual-luciferase gene expression in mouse
10.3760/cma.j.issn.1003-9279.2012.06.012
- VernacularTitle:重组8型腺相关病毒介导双荧光素酶基因在小鼠体内的表达
- Author:
Gang WANG
1
;
CHI-Jie YU
;
Xiao-Yan DONG
;
Wen-Hong TIAN
;
Xiao-Bing WU
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Dependovirus;
Lueiferase;
Expressed sequence tags
- From:
Chinese Journal of Experimental and Clinical Virology
2012;26(6):439-441
- CountryChina
- Language:Chinese
-
Abstract:
Objective Recombinant adeno-associated virus type 8 (rAAV8) mediating transgene expression in mice was investigated using co-expressed report gene of secreted Gaussia princeps luciferase (Gluc) and non-secreted firefly luciferase(Fluc).Methods rAAV8-Gluc/Fluc was prepared and infected HEK293 cells to test its performance in vitro.BALB/c mice were received rAAV8-Gluc/Fluc at different doses by intravenous injection (iv) or intramuscular injection (im).Then Gluc activities in blood were measured,the whole-body images for Fluc activities were performed and Fluc activities of tissue lysate were also detected.Results rAAV8-Gluc/Fluc was successfully prepared and could infected HEK293 cells.The Gluc was mainly detected in the culture media while the Fluc was mainly detected within cells.The blood Gluc activities of mice received rAAV8-Gluc/Fluc by iv or im peaked at 10-20 d post injection and persisted for at least 120d.The blood Gluc activities increased at the rAAV8-Gluc/Fluc dose-dependent manner.For mice received rAAV8 by iv,Fluc mainly expressed in liver and minor Fluc expression was also found in cardiac muscle and skeletal muscle.For mice received rAAV8-Gluc/Fluc by im,Fluc mainly expressed in local skeletal muscle and secondly in liver.Conclusion rAAV8-Gluc/Fluc has been prepared successfully and its mediating transgene expression in mice has been investigated.This research will facilitate preclinical studies for rAAV8-mediated gene therapy.
