title: Analysis of mutations within the intron20 splice donor site of CREBBP in patients with and without classical RSTS.
abstract: Whole-exome sequencing of a patient with intellectual disability and without recognisable phenotype yielded a mutation in the intron20 splice donor site of CREBBP. Mutations at different positions within the same intron20 splice donor site were observed in three patients clinically suspected as having Rubinstein-Taybi syndrome (RSTS). All mutations were de novo and likely disease-causing. To investigate a putative difference in splicing between the patient without RSTS phenotype and the three patients with the RSTS phenotype, we analysed the effects of these mutations on splicing of the pre-mRNA of CREBBP. As no RNA of patients was available, we generated a new and improved exon-trap vector, pCDNAGHE, and tested the effect of the various mutations on splicing in vitro. All mutations lead to skipping of exon20. In one of the patients with an RSTS phenotype, there was also some normal splicing detectable. We conclude that the splicing pattern obtained by exon-trapping cannot explain the difference in phenotype between the patient without the RSTS phenotype and the patients with clinical RSTS. Patient or tissue-specific splice effects as well as modifying genes likely will explain the difference in phenotype.
authors.affiliation: Department of Human Genetics, Leiden University Medical Centre, Leiden, The Netherlands.|Department of Clinical Genetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Johannes G
authors.initials: JG
authors.lastname: Dauwerse
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Martine
authors.initials: M
authors.lastname: van Belzen
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Arie
authors.initials: A
authors.lastname: van Haeringen
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Gijs
authors.initials: G
authors.lastname: van Santen
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Christian
authors.initials: C
authors.lastname: van de Lans
authors.affiliation: PEDEGO Research Unit, Department of Clinical Genetics, University of Oulu, MRC Oulu, Oulu University Hospital, Oulu, Finland.
authors.forename: Elisa
authors.initials: E
authors.lastname: Rahikkala
authors.affiliation: Genetics Unit, Department of Obstetrics and Paediatrics, IRCCS S. Maria Nuova Hospital, Reggio Emilia, Italy.
authors.forename: Livia
authors.initials: L
authors.lastname: Garavelli
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Martijn
authors.initials: M
authors.lastname: Breuning
authors.affiliation: Department of Paediatrics, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Raoul
authors.initials: R
authors.lastname: Hennekam
authors.affiliation: Department of Human Genetics, Leiden University Medical Centre, Leiden, The Netherlands.
authors.forename: Dorien
authors.initials: D
authors.lastname: Peters
mesh_terms: D000818:Animals; D050882:CREB-Binding Protein / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002648:Child; D002675:Child, Preschool; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D006801:Humans; D007438:Introns; D008297:Male; D009154:Mutation; D010641:Phenotype; D012326:RNA Splicing; D012415:Rubinstein-Taybi Syndrome / Q000175:diagnosis / Q000235:genetics*
chemical_list: D050882:CREB-Binding Protein; C494363:CREBBP protein, human