title: A simple and efficient purification of transduced cells by using green fluorescent protein gene as a selection marker.
abstract: BACKGROUND
  Simple and efficient method for the selection of transduced cells would greatly facilitate the clinical utilization of retrovirus vectors. We developed a therapeutic bicistronic retrovirus vector for Gaucher disease, MFG-GC-GFP, which contains the human glucocerebrosidase (GC) gene and the green fluorescent protein (GFP) gene of the jellyfish Aequorea victoria as a vital selection marker, and investigated its applicability as gene therapy for Gaucher disease.
  
  
  RESULTS
  A packaging cell line, GP + envAM12, was transfected with MFG-GC-GFP and, thus, produced a high titer recombinant virus (1.0 x 10(6) c.f.u./mL) in the culture supernatant. The expression level of GFP was correlated with the virus production in cells. The recombinant virus infected skin fibroblasts from a Gaucher patient and a sorted fraction of the cells expressing GFP by flow cytometry exhibited almost a six-fold higher activity of GC than normal fibroblasts.
  
  
  CONCLUSIONS
  These data indicate that MFG-GC-GFP enables the one-step purification of a transduced fraction of target cells and is, therefore, considered to be a useful therapeutic vector for the experimental gene therapy of Gaucher disease.
authors.affiliation: Department of Pediatrics, Tokai University School of Medicine, Kanagawa, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Shimizu
authors.forename: K
authors.initials: K
authors.lastname: Ando
authors.forename: M
authors.initials: M
authors.lastname: Kimura
authors.forename: H
authors.initials: H
authors.lastname: Miyatake
authors.forename: S
authors.initials: S
authors.lastname: Inokuchi
authors.forename: I
authors.initials: I
authors.lastname: Takakura
authors.forename: M
authors.initials: M
authors.lastname: Migita
authors.forename: T
authors.initials: T
authors.lastname: Shimada
authors.forename: S
authors.initials: S
authors.lastname: Kato
mesh_terms: D000818:Animals; D002460:Cell Line; D005776:Gaucher Disease / Q000628:therapy; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D005822:Genetic Vectors; D005962:Glucosylceramidase / Q000235:genetics; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics*; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D005819:Genetic Markers; D008164:Luminescent Proteins; D005962:Glucosylceramidase