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

Brown, Fiona C.; Scott, Nicholas A.; Rank, Gerhard; Collinge, J; Vadolas, Jim; Vickaryous, Nicola; Whitelaw, Nadia C; Whitelaw, Emma; Kile, Benjamin T.; Jane, Stephen M.; Curtis, David J.

doi:10.1016/j.bcmd.2012.09.004

Cited by 13 publications

(22 citation statements)

References 38 publications

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“…A third line, MommeD7 , also displayed high mean fluorescence, as was previously noted (Ashe et al 2008). Analysis of MommeD7 +/− blood samples revealed a high reticulocyte count and a mutation was subsequently identified in the canonical polyadenylation signal of the haemoglobin, beta major gene (Brown et al 2013). Full blood count analysis and reticulocyte counts of MommeD6 +/− mice showed no differences from wild-type littermates (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…For example, mutations in genes alter the red blood cell count or affect stability of the GFP RNA or protein (Garneau et al 2007). For instance, the high mean fluorescence per expressing cell observed in MommeD7 results from a mutation in the polyadenylation signal of the haemoglobin, beta adult major chain gene which leads to low mean corpuscular volume and reticulocytosis (Brown et al 2013). MommeR1 , a mutant line identified from a modified screen to identify recessive mutants, also displays an unusually high mean fluorescence per expressing cell compared to shift in the percentage of GFP expressing cells.…”

Section: Discussionmentioning

confidence: 99%

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The first mouse mutants of D14Abb1e (Fam208a) show that it is critical for early development

et al. 2014

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An ENU mutagenesis screen to identify novel epigenetic modifiers was established in mice carrying a multi-copy GFP transgene, which is expressed in a variegated manner in erythrocytes and is highly sensitive to epigenetic silencing. The screen has produced mouse mutants of both known modifiers of epigenetic state, such as Dnmt1 and Smarca5, and novel modifiers, such as Smchd1 and Rlf. Here we report two mouse lines generated from the screen, MommeD6 and MommeD20, with point mutations in D14Abb1e. These are the first mouse mutants of D14Abb1e (alsoknownasFam208a), a gene about which little is known. Heterozygous intercrosses show that homozygous mutants from both the MommeD6 and MommeD20 lines are not viable beyond gastrulation, demonstrating an important role for D14Abb1e in development. We demonstrate that haploinsufficiency for D14Abb1e effects transgene expression at the RNA level. Analysis of the predicted D14Abb1e protein sequence reveals that it contains putative nuclear localisation signals and a domain of unknown function, DUF3715. Our studies reveal that D14Abb1e is localised to the nucleus and is expressed in skin and testes.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The first mouse mutants of D14Abb1e (Fam208a) show that it is critical for early development

et al. 2014

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“…In an ENU mutagenesis screen to identify genes regulating erythropoiesis (Rank et al, 2009;Brown et al, 2013Brown et al, , 2015, a G 1 mouse (RBC12) was identified with a mean cell volume (MCV) that was more than three standard deviations lower than that of the overall G 1 population (MCV 38 fl vs. 44Á9 fl). When the founder animal was mated with wild-type C57BL/6, c. 50% of pups born exhibited a reduced MCV, indicating that the phenotype was heritable, dominant and highly penetrant.…”

Section: Identification Of a Novel Mouse Model Of Microcytic Anaemiamentioning

confidence: 99%

Loss of Dynamin 2 GTPase function results in microcytic anaemia

et al. 2017

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In a dominant mouse ethylnitrosurea mutagenesis screen for genes regulating erythropoiesis, we identified a pedigree with a novel microcytic hypochromia caused by a V235G missense mutation in Dynamin 2 (Dnm2). Mutations in Dnm2, a GTPase, are highly disease-specific and have been implicated in four forms of human diseases: centronuclear myopathy, Charcot-Marie Tooth neuropathy and, more recently, T-cell leukaemia and Hereditary Spastic Paraplegia, but red cell abnormalities have not been reported to date. The V235G mutation lies within a crucial GTP nucleotide-binding pocket of Dnm2, and resulted in defective GTPase activity and incompatibility with life in the homozygous state. Dnm2 is an essential mediator of clathrin-mediated endocytosis, which is required for the uptake of transferrin (Tf) into red cells for incorporation of haem. Accordingly, we observed significantly reduced Tf uptake by Dnm2 cells, which led to impaired endosome formation. Despite these deficiencies, surprisingly all iron studies were unchanged, suggesting an unexplained alternative mechanism underlies microcytic anaemia in Dnm2 mice. This study provides the first in vivo evidence for the requirements of Dnm2 in normal erythropoiesis.

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“…Using ENU mutagenesis to discover new alleles that regulate erythropoiesis, 15 we identified a mouse (RBC10) in the first generation (G1) with a mean cell volume (MCV) .3 standard deviations (SDs) below the normal (MCV, 39 vs 45 fl in wild-type mice). Fifty percent of pups born from affected mice crossed with wild-type mice displayed a reduced MCV, indicating that the phenotype was fully penetrant (data not shown).…”

Section: Characterization Of a Mouse Line With Microcytic Anemiamentioning

confidence: 99%

“…15,16 KCC1 M935K mice were maintained on a mixed C57BL/6J 3 BALB/c background because the microcytic phenotype was less severe when backcrossed onto a pure C57BL/6J background. Mice were genotyped by polymerase chain reaction (PCR) amplification of the region spanning the M935K mutation using primers: forward (59-AGCTGAAGTG GAGGTGGTAGAGAT-39) and reverse (59-TCCAGCCTAAGAGCCGAG TG-39), followed by sequencing the PCR product using the Big Dye Terminator reagents (Australian Genome Research Facility) to identify the point mutation.…”

Section: M935kmentioning

confidence: 99%

Activation of the erythroid K-Cl cotransporter Kcc1 enhances sickle cell disease pathology in a humanized mouse model

Brown

Conway

Cerruti

et al. 2015

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

Cited by 13 publications

References 38 publications

The first mouse mutants of D14Abb1e (Fam208a) show that it is critical for early development

The first mouse mutants of D14Abb1e (Fam208a) show that it is critical for early development

Loss of Dynamin 2 GTPase function results in microcytic anaemia

Activation of the erythroid K-Cl cotransporter Kcc1 enhances sickle cell disease pathology in a humanized mouse model

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