title: Humanized beta-thalassemia mouse model containing the common IVSI-110 splicing mutation.
abstract: Splicing mutations are common causes of beta-thalassemia. Some splicing mutations permit normal splicing as well as aberrant splicing, which can give a reduced level of normal beta-globin synthesis causing mild disease (thalassemia intermedia). For other mutations, normal splicing is reduced to low levels, and patients are transfusion-dependent when homozygous for the disease. The development of therapies for beta-thalassemia will require suitable mouse models for preclinical studies. In this study, we report the generation of a humanized mouse model carrying the common IVSI-110 splicing mutation on a BAC including the human beta-globin ((hu)beta-globin) locus. We examined heterozygous murine beta-globin knock-out mice ((mu)beta(th-3/+)) carrying either the IVSI-110 or the normal (hu)beta-globin locus. Our results show a 90% decrease in (hu)beta-globin chain synthesis in the IVSI-110 mouse model compared with the mouse model carrying the normal (hu)beta-globin locus. This notable difference is attributed to aberrant splicing. The humanized IVSI-110 mouse model accurately recapitulates the splicing defect found in comparable beta-thalassemia patients. This mouse model is available as a platform for testing strategies for the restoration of normal splicing.
authors.affiliation: Cell and Gene Therapy Research Group, Murdoch Childrens Research Institute, The University of Melbourne, Royal Children's Hospital, Parkville 3052, Melbourne, Australia. jim.vadolas@mcri.edu.au
authors.forename: Jim
authors.initials: J
authors.lastname: Vadolas
authors.forename: Mikhail
authors.initials: M
authors.lastname: Nefedov
authors.forename: Hady
authors.initials: H
authors.lastname: Wardan
authors.forename: Sima
authors.initials: S
authors.lastname: Mansooriderakshan
authors.forename: Lucille
authors.initials: L
authors.lastname: Voullaire
authors.forename: Duangporn
authors.initials: D
authors.lastname: Jamsai
authors.forename: Robert
authors.initials: R
authors.lastname: Williamson
authors.forename: Panayiotis A
authors.initials: PA
authors.lastname: Ioannou
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004195:Disease Models, Animal; D005816:Genetic Complementation Test; D005838:Genotype; D005914:Globins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes; D017086:beta-Thalassemia / Q000235:genetics*
chemical_list: D006454:Hemoglobins; D005914:Globins