title: Correction of the Pathogenic Alternative Splicing, Caused by the Common GNB3 c.825C>T Allele, Using a Novel, Antisense Morpholino.
abstract: The very common GNB3 c.825C>T polymorphism (rs5443) is present in approximately half of all human chromosomes. Significantly, the presence of the GNB3 825T allele has been strongly associated with predisposition to essential hypertension. Paradoxically the presence of the GNB3 825T allele, in exon 10, introduces a pathogenic alternative RNA splice site into the middle of exon 9. To attempt to correct this pathogenic aberrant splicing, we, therefore, bioinformatically designed, using a Gene Tools(®) algorithm, a GNB3-specific, antisense morpholino. It was hoped that this morpholino would behave in vitro as either a potential splice blocker and/or exon skipper, to both bind and inhibit/reduce the aberrant splicing of the GNB3 825T allele. On transfecting a human lymphoblast cell line homozygous for the 825T allele, with this antisense morpholino, we encouragingly observed both a significant reduction (from ∼58% to ∼5%) in the production of the aberrant smaller GNB3 transcript, and a subsequent increase in the normal GNB3 transcript (from ∼42% to ∼95%). Our results demonstrate the potential use of a GNB3-specific antisense morpholino, as a pharmacogenetic therapy for essential hypertension.
authors.affiliation: 1 School of Science, Engineering & Technology, Abertay University , Dundee, United Kingdom .|2 Blood Sciences Laboratory, Department of Haematology, Ninewells Hospital , Dundee, United Kingdom .
authors.forename: Jonathan C P
authors.initials: JC
authors.lastname: McGlinchey
authors.affiliation: 3 Centre for Paediatrics, Barts and The London School of Medicine and Dentistry, Queen Mary University of London , Barts and The London Children's Hospital, London, United Kingdom .
authors.forename: Hemanth
authors.initials: H
authors.lastname: Tummala
authors.affiliation: 1 School of Science, Engineering & Technology, Abertay University , Dundee, United Kingdom .
authors.forename: Douglas H
authors.initials: DH
authors.lastname: Lester
mesh_terms: D000483:Alleles; D017398:Alternative Splicing / Q000235:genetics; D002460:Cell Line; D000075222:Essential Hypertension / Q000235:genetics / Q000628:therapy*; D020022:Genetic Predisposition to Disease; D020962:Heterotrimeric GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D006801:Humans; D060172:Morpholinos / Q000235:genetics* / Q000627:therapeutic use; D020320:Oligoribonucleotides, Antisense / Q000235:genetics* / Q000627:therapeutic use; D022821:RNA Splice Sites / Q000235:genetics
chemical_list: C454420:G-protein beta3 subunit; D060172:Morpholinos; D020320:Oligoribonucleotides, Antisense; D022821:RNA Splice Sites; D020962:Heterotrimeric GTP-Binding Proteins