title: GALNS gene expression profiling in Morquio A patients' fibroblasts.
abstract: BACKGROUND
  Quantification studies of mutated mRNAs have not been carried out on Morquio A patients. Such studies are very important for the determination of stability of premature termination codons (PTC) bearing transcripts in order to assess the appropriateness of introducing the newly developed therapeutic strategies such as "stop codon read-through therapy".
  
  
  METHODS
  This paper focuses on the study of the GALNS gene and mRNAs in two severe forms of Morquio A patients' fibroblasts with development of a new and rapid real-time RT-PCR for detection and quantification of absolute mRNA copy number.
  
  
  RESULTS
  We identified two new mutations c.385A>T (p.K129X) and c.899-1G>C) in Pt1 and a known splicing defect c.120+1G>A in Pt2. Using RT-PCR and real-time RT-PCR in Pt2 we detected low levels of mRNAs, suggesting its instability; in Pt1, we detected three aberrant mRNAs introducing premature stop codons, suggesting that both the c.385A>T and c.899-1G>C mutations produce mRNAs capable of escaping the nonsense-mediated decay (NMD) pathway.
  
  
  CONCLUSIONS
  The development of a real-time RT-PCR assay allows to absolutely quantify the GALNS mRNAs carrying mutations that lead to PTCs bearing transcripts, which escape the NMD process and are potentially suitable for the new therapeutic approach.
authors: Metabolic and Muscular Unit, Clinic of Pediatric Neurology, AOU Meyer, Florence, Italy.
authors: L
authors: L
authors: Carraresi
authors: R
authors: R
authors: Parini
authors: C
authors: C
authors: Filoni
authors: A
authors: A
authors: Caciotti
authors: G
authors: G
authors: Sersale
authors: S
authors: S
authors: Tomatsu
authors: C
authors: C
authors: Orlando
authors: E
authors: E
authors: Zammarchi
authors: R
authors: R
authors: Guerrini
authors: M A
authors: MA
authors: Donati
authors: A
authors: A
authors: Morrone
mesh_terms: D000483:Alleles; D002648:Child; D002811:Chondroitinsulfatases / Q000235:genetics*; D018389:Codon, Nonsense / Q000235:genetics; D005091:Exons / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000201:enzymology*; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D009085:Mucopolysaccharidosis IV / Q000235:genetics*; D009154:Mutation; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D018389:Codon, Nonsense; D012333:RNA, Messenger; D002811:Chondroitinsulfatases; C102295:GALNS protein, human