ENU mutagenesis identifies the first mouse mutants reproducing human beta-thalassemia at the genomic level

Fiona Brown, Nicholas Scott, Gerhard Rank, Janelle E Collinge, Jim Vadolas, Nicola Vickaryous, Nadia C Whitelaw, Emma Whitelaw, Benjamin Kile, Stephen Marsden Jane, David John Curtis

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Forward genetic screens have been performed in many species to identify phenotypes in specific organ systems. We have undertaken a large-scale N-ethyl-N-nitrosourea (ENU) mutagenesis screen to identify dominant mutations that perturb erythropoiesis in mice. Mutant mice that displayed an erythrocyte mean cell volume (MCV) greater than three standard deviations from the population mean were identified. Two of these lines, RBC13 and RBC14, displayed a hypochromic, microcytic anemia, accompanied by a marked reticulocytosis, splenomegaly and diminished red cell survival. Timed pregnancies from heterozygous intercrosses revealed that a quarter of the embryos displayed severe anemia and did not survive beyond embryonic day (E) 18.5, consistent with homozygous ?-thalassemia. Genetic complementation studies with a ?-thalassemia mouse line reproduced the embryonic lethality in compound heterozygotes and a genomic custom capture array and massively parallel sequencing of the ?-globin locus identified the causative mutations. The RBC13 line displayed a nonsense mutation at codon 40 in exon 2 of the ?-major gene, invoking parallels with the common ?039 thalassemia mutation seen in humans. The RBC14 line exhibited a mutation at the polyadenylation signal of the ?-major gene, exactly replicating a human ?-thalassemia mutation. The RBC13 and RBC14 lines are the first ?-thalassemia mouse models that reproduce human ?-thalassemia at the genomic level, and as such highlight the power of ENU mutagenesis screens in generating mouse models of human disease. ? 2012 Elsevier Inc..
Original languageEnglish
Pages (from-to)86-92
Number of pages7
JournalBlood Cells, Molecules, and Diseases
Volume50
Issue number2
DOIs
Publication statusPublished - 2013

Cite this

Brown, Fiona ; Scott, Nicholas ; Rank, Gerhard ; Collinge, Janelle E ; Vadolas, Jim ; Vickaryous, Nicola ; Whitelaw, Nadia C ; Whitelaw, Emma ; Kile, Benjamin ; Jane, Stephen Marsden ; Curtis, David John. / ENU mutagenesis identifies the first mouse mutants reproducing human beta-thalassemia at the genomic level. In: Blood Cells, Molecules, and Diseases. 2013 ; Vol. 50, No. 2. pp. 86-92.
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abstract = "Forward genetic screens have been performed in many species to identify phenotypes in specific organ systems. We have undertaken a large-scale N-ethyl-N-nitrosourea (ENU) mutagenesis screen to identify dominant mutations that perturb erythropoiesis in mice. Mutant mice that displayed an erythrocyte mean cell volume (MCV) greater than three standard deviations from the population mean were identified. Two of these lines, RBC13 and RBC14, displayed a hypochromic, microcytic anemia, accompanied by a marked reticulocytosis, splenomegaly and diminished red cell survival. Timed pregnancies from heterozygous intercrosses revealed that a quarter of the embryos displayed severe anemia and did not survive beyond embryonic day (E) 18.5, consistent with homozygous ?-thalassemia. Genetic complementation studies with a ?-thalassemia mouse line reproduced the embryonic lethality in compound heterozygotes and a genomic custom capture array and massively parallel sequencing of the ?-globin locus identified the causative mutations. The RBC13 line displayed a nonsense mutation at codon 40 in exon 2 of the ?-major gene, invoking parallels with the common ?039 thalassemia mutation seen in humans. The RBC14 line exhibited a mutation at the polyadenylation signal of the ?-major gene, exactly replicating a human ?-thalassemia mutation. The RBC13 and RBC14 lines are the first ?-thalassemia mouse models that reproduce human ?-thalassemia at the genomic level, and as such highlight the power of ENU mutagenesis screens in generating mouse models of human disease. ? 2012 Elsevier Inc..",
author = "Fiona Brown and Nicholas Scott and Gerhard Rank and Collinge, {Janelle E} and Jim Vadolas and Nicola Vickaryous and Whitelaw, {Nadia C} and Emma Whitelaw and Benjamin Kile and Jane, {Stephen Marsden} and Curtis, {David John}",
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ENU mutagenesis identifies the first mouse mutants reproducing human beta-thalassemia at the genomic level. / Brown, Fiona; Scott, Nicholas; Rank, Gerhard; Collinge, Janelle E; Vadolas, Jim; Vickaryous, Nicola; Whitelaw, Nadia C; Whitelaw, Emma; Kile, Benjamin; Jane, Stephen Marsden; Curtis, David John.

In: Blood Cells, Molecules, and Diseases, Vol. 50, No. 2, 2013, p. 86-92.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - ENU mutagenesis identifies the first mouse mutants reproducing human beta-thalassemia at the genomic level

AU - Brown, Fiona

AU - Scott, Nicholas

AU - Rank, Gerhard

AU - Collinge, Janelle E

AU - Vadolas, Jim

AU - Vickaryous, Nicola

AU - Whitelaw, Nadia C

AU - Whitelaw, Emma

AU - Kile, Benjamin

AU - Jane, Stephen Marsden

AU - Curtis, David John

PY - 2013

Y1 - 2013

N2 - Forward genetic screens have been performed in many species to identify phenotypes in specific organ systems. We have undertaken a large-scale N-ethyl-N-nitrosourea (ENU) mutagenesis screen to identify dominant mutations that perturb erythropoiesis in mice. Mutant mice that displayed an erythrocyte mean cell volume (MCV) greater than three standard deviations from the population mean were identified. Two of these lines, RBC13 and RBC14, displayed a hypochromic, microcytic anemia, accompanied by a marked reticulocytosis, splenomegaly and diminished red cell survival. Timed pregnancies from heterozygous intercrosses revealed that a quarter of the embryos displayed severe anemia and did not survive beyond embryonic day (E) 18.5, consistent with homozygous ?-thalassemia. Genetic complementation studies with a ?-thalassemia mouse line reproduced the embryonic lethality in compound heterozygotes and a genomic custom capture array and massively parallel sequencing of the ?-globin locus identified the causative mutations. The RBC13 line displayed a nonsense mutation at codon 40 in exon 2 of the ?-major gene, invoking parallels with the common ?039 thalassemia mutation seen in humans. The RBC14 line exhibited a mutation at the polyadenylation signal of the ?-major gene, exactly replicating a human ?-thalassemia mutation. The RBC13 and RBC14 lines are the first ?-thalassemia mouse models that reproduce human ?-thalassemia at the genomic level, and as such highlight the power of ENU mutagenesis screens in generating mouse models of human disease. ? 2012 Elsevier Inc..

AB - Forward genetic screens have been performed in many species to identify phenotypes in specific organ systems. We have undertaken a large-scale N-ethyl-N-nitrosourea (ENU) mutagenesis screen to identify dominant mutations that perturb erythropoiesis in mice. Mutant mice that displayed an erythrocyte mean cell volume (MCV) greater than three standard deviations from the population mean were identified. Two of these lines, RBC13 and RBC14, displayed a hypochromic, microcytic anemia, accompanied by a marked reticulocytosis, splenomegaly and diminished red cell survival. Timed pregnancies from heterozygous intercrosses revealed that a quarter of the embryos displayed severe anemia and did not survive beyond embryonic day (E) 18.5, consistent with homozygous ?-thalassemia. Genetic complementation studies with a ?-thalassemia mouse line reproduced the embryonic lethality in compound heterozygotes and a genomic custom capture array and massively parallel sequencing of the ?-globin locus identified the causative mutations. The RBC13 line displayed a nonsense mutation at codon 40 in exon 2 of the ?-major gene, invoking parallels with the common ?039 thalassemia mutation seen in humans. The RBC14 line exhibited a mutation at the polyadenylation signal of the ?-major gene, exactly replicating a human ?-thalassemia mutation. The RBC13 and RBC14 lines are the first ?-thalassemia mouse models that reproduce human ?-thalassemia at the genomic level, and as such highlight the power of ENU mutagenesis screens in generating mouse models of human disease. ? 2012 Elsevier Inc..

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