title: Knock-out mouse for Canavan disease: a model for gene transfer to the central nervous system.
abstract: BACKGROUND
  Canavan disease (CD) is an autosomal recessive leukodystrophy characterized by deficiency of aspartoacylase (ASPA) and increased levels of N-acetylaspartic acid (NAA) in brain and body fluids, severe mental retardation and early death. Gene therapy has been attempted in a number of children with CD. The lack of an animal model has been a limiting factor in developing vectors for the treatment of CD. This paper reports the successful creation of a knock-out mouse for Canavan disease that can be used for gene transfer.
  
  
  METHODS
  Genomic library lambda knock-out shuttle (lambdaKOS) was screened and a specific pKOS/Aspa clone was isolated and used to create a plasmid with 10 base pair (bp) deletion of exon four of the murine aspa. Following linearization, the plasmid was electroporated to ES cells. Correctly targeted ES clones were identified following positive and negative selection and confirmed by Southern analysis. Chimeras were generated by injection of ES cells to blastocysts. Germ line transmission was achieved by the birth of heterozygous mice as confirmed by Southern analysis.
  
  
  RESULTS
  Heterozygous mice born following these experiments have no overt phenotype. The homozygous mice display neurological impairment, macrocephaly, generalized white matter disease, deficient ASPA activity and high levels of NAA in urine. Magnetic resonance imaging (MRI) and spectroscopy (MRS) of the brain of the homozygous mice show white matter changes characteristic of Canavan disease and elevated NAA levels.
  
  
  CONCLUSIONS
  The newly created ASPA deficient mouse establishes an important animal model of Canavan disease. This model should be useful for developing gene transfer vectors to treat Canavan disease. Vectors for the central nervous system (CNS) and modulation of NAA levels in the brain should further add to the understanding of the pathophysiology of Canavan disease. Data generated from this animal model will be useful for developing strategies for gene therapy in other neurodegenerative diseases.
authors.affiliation: Department of Pediatrics, Children's Hospital, UTMB Galveston, TX 77555-0359, USA. rmatalon@utmb.edu
authors.forename: R
authors.initials: R
authors.lastname: Matalon
authors.forename: P L
authors.initials: PL
authors.lastname: Rady
authors.forename: K A
authors.initials: KA
authors.lastname: Platt
authors.forename: H B
authors.initials: HB
authors.lastname: Skinner
authors.forename: M J
authors.initials: MJ
authors.lastname: Quast
authors.forename: G A
authors.initials: GA
authors.lastname: Campbell
authors.forename: K
authors.initials: K
authors.lastname: Matalon
authors.forename: J D
authors.initials: JD
authors.lastname: Ceci
authors.forename: S K
authors.initials: SK
authors.lastname: Tyring
authors.forename: M
authors.initials: M
authors.lastname: Nehls
authors.forename: S
authors.initials: S
authors.lastname: Surendran
authors.forename: J
authors.initials: J
authors.lastname: Wei
authors.forename: E L
authors.initials: EL
authors.lastname: Ezell
authors.forename: S
authors.initials: S
authors.lastname: Szucs
mesh_terms: D000581:Amidohydrolases / Q000235:genetics; D000818:Animals; D001921:Brain / Q000002:abnormalities / Q000473:pathology; D017825:Canavan Disease / Q000235:genetics* / Q000628:therapy; D002490:Central Nervous System / Q000378:metabolism*; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D008279:Magnetic Resonance Imaging; D051379:Mice; D018345:Mice, Knockout; D010641:Phenotype
chemical_list: D000581:Amidohydrolases; C019751:aspartoacylase