title: Sleeping Beauty transposon system is a reliable gene delivery tool for hereditary tyrosinaemia type 1 disease gene therapy: size of the foreign gene decides the timing of stable integration into the host chromosomes.
abstract: OBJECTIVE
  This study investigated compensation for loss of the fumaryl-acetoacetate hydrolase gene (Fah) by gene therapy using the Sleeping Beauty transposon system (SBTS), in a hereditary tyrosinaemia type 1 (HT-1) mouse model (Fah-/-).
  
  
  METHODS
  Twenty Fah-/-  study mice, five wild-type positive controls and five Fah-/- negative controls were included. All Fah-/- mice received 2-(2-nitro-4-trifluoro-methylbenzoyl)-1,3-cyclo hexaedione (NTBC). Fah-/- study mice were randomly injected with one of two  SBTS constructs: Fah-SBTS (containing mouse Fah gene), or forkhead box M1b (FOXM1B)-Fah-SBTS (containing mouse Fah and human FOXM1B genes). Firefly luciferase-SBTS was injected as a trace marker. NTBC treatment stopped after construct injection;  Fah-/- negative controls were kept healthy with continued NTBC. Mice were weighed daily; the luciferase signal was monitored by in vivo bioluminescence, and Fah and FOXM1B gene expression were evaluated.
  
  
  RESULTS
  The Fah gene integrated into the mouse  chromosomes within 1 week of Fah-SBTS injection (mice survived without NTBC thereafter) and within 1 month of FOXM1B-Fah-SBTS injection (mice lost weight dramatically and needed additional NTBC).
  
  
  CONCLUSIONS
  The shorter Fah gene had an advantage over the longer FOXM1B-Fah  gene for stable integration into the host mouse chromosomes.
authors.affiliation: Stem Cell Research Institute, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: X-J
authors.initials: XJ
authors.lastname: Pan
authors.forename: Z-Z
authors.initials: ZZ
authors.lastname: Ma
authors.forename: Q-J
authors.initials: QJ
authors.lastname: Zhang
authors.forename: L
authors.initials: L
authors.lastname: Fan
authors.forename: Q-H
authors.initials: QH
authors.lastname: Li
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D033481:Chromosomes, Mammalian / Q000235:genetics*; D004251:DNA Transposable Elements; D005260:Female; D000072278:Forkhead Box Protein M1; D051858:Forkhead Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy; D006801:Humans; D006867:Hydrolases / Q000096:biosynthesis / Q000235:genetics; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D018345:Mice, Knockout; D016254:Mutagenesis, Insertional; D020176:Tyrosinemias / Q000235:genetics / Q000628:therapy*
chemical_list: D004251:DNA Transposable Elements; C107585:FOXM1 protein, human; D000072278:Forkhead Box Protein M1; D051858:Forkhead Transcription Factors; D049409:Luciferases, Firefly; D006867:Hydrolases; C020156:fumarylacetoacetase