title: [Recombinant adeno-associated virus type 8 mediated dual-luciferase gene expression in mouse].
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 Glue activities in blood were measured,the whole-body images for Flue activities were performed and Flue activities of tissue lysate were also detected.
  
  
  RESULTS
  rAAV8-Gluc/Fluc was successfully prepared and could infected HEK293 cells. The Glue was mainly detected in the culture media while the Flue was mainly detected within cells. The blood Glue 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 1000052, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Chi-Jie
authors.initials: CJ
authors.lastname: Yu
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Dong
authors.forename: Wen-Hong
authors.initials: WH
authors.lastname: Tian
authors.forename: Xiao-Bing
authors.initials: XB
authors.lastname: Wu
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000295:instrumentation; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008099:Liver / Q000201:enzymology; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000201:enzymology
chemical_list: D008156:Luciferases