title: Multiple abnormally spliced ABCA1 mRNAs caused by a novel splice site mutation of ABCA1 gene in a patient with Tangier disease.
abstract: BACKGROUND
  Mutations in ABCA1 gene are the cause of Tangier disease (TD) and familial high density lipoprotein (HDL) deficiency. Splice site mutations of this gene were reported infrequently.
  
  
  METHODS
  ABCA1 gene was sequenced in a TD patient and in subjects with low HDL. The effect of intronic variants on ABCA1 pre-mRNA splicing was studied in COS-1 cells expressing a mutant minigene or in patients' cells.
  
  
  RESULTS
  A novel mutation in intron 20 (c.2961 -2 A>C) was found in the TD patient. To assess its effect, a mutant ABCA1 minigene, containing intron 18-intron 23 region, was expressed in COS-1 cells. The mutant minigene generated three transcripts: i) in the first (459bp) 61 nucleotides of intron 20 were retained; ii) in the second (384bp) exon 20 joined to exon 21 devoid of the first 14 nucleotides; and iii) in the third (255bp) the entire exon 21 was skipped. The first two transcripts were also observed in patient's peripheral blood mononuclear cells. These mRNAs encode truncated proteins. A variant in intron 8 (c.814 -14 ins A), identified in subjects with low HDL, had no effect on ABCA1 pre-mRNA splicing.
  
  
  CONCLUSIONS
  Functional analysis is required to establish the effect of intronic mutations on ABCA1 pre-mRNA splicing.
authors.affiliation: Department of Biomedical Sciences, University of Modena and Reggio Emilia, Italy.
authors.forename: Letizia
authors.initials: L
authors.lastname: Bocchi
authors.forename: Livia
authors.initials: L
authors.lastname: Pisciotta
authors.forename: Tommaso
authors.initials: T
authors.lastname: Fasano
authors.forename: Chiara
authors.initials: C
authors.lastname: Candini
authors.forename: Maria Rita
authors.initials: MR
authors.lastname: Puntoni
authors.forename: Tiziana
authors.initials: T
authors.lastname: Sampietro
authors.forename: Stefano
authors.initials: S
authors.lastname: Bertolini
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Calandra
mesh_terms: D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000328:Adult; D006801:Humans; D008297:Male; D009154:Mutation; D022821:RNA Splice Sites / Q000235:genetics*; D012326:RNA Splicing / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D013631:Tangier Disease / Q000235:genetics*
chemical_list: C577836:ABCA1 protein, human; D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters; D022821:RNA Splice Sites; D012333:RNA, Messenger