Open Access Research

An ENU mutagenesis screen identifies novel and known genes involved in epigenetic processes in the mouse

Lucia Daxinger12, Sarah K Harten1, Harald Oey12, Trevor Epp16, Luke Isbel12, Edward Huang1, Nadia Whitelaw1, Anwyn Apedaile1, Anabel Sorolla1, Joan Yong1, Vandhana Bharti1, Joanne Sutton1, Alyson Ashe17, Zhenyi Pang1, Nathan Wallace1, Daniel J Gerhardt3, Marnie E Blewitt45, Jeffrey A Jeddeloh3 and Emma Whitelaw12*

Author Affiliations

1 Epigenetics Laboratory, QIMR Berghofer Medical Research Institute, Herston, Qld 4006, Australia

2 La Trobe Institute for Molecular Science, Department of Genetics, La Trobe University, Bundoora 3086, Vic, Australia

3 Development and Research, Roche NimbleGen, 500 South Rosa Road, Madison, WI 53705, USA

4 Molecular Medicine Division, The Walter and Eliza Hall Institute of Medical Research, University of Melbourne, Melbourne 3050, Vic, Australia

5 Department of Medical Biology and Dept of Genetics, University of Melbourne, Melbourne 3050, Vic, Australia

6 Present address: Institute of Molecular Genetics of ASCR, Videnska 1083, Prague 4, Czech Republic

7 Present address: Gurdon Institute, University of Cambridge, Cambridge CB2 IQN, UK

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Genome Biology 2013, 14:R96  doi:10.1186/gb-2013-14-9-r96

Published: 11 September 2013

Additional files

Additional file 1:

Linked intervals. Manhattan plots showing linked intervals identified by Illumina GoldenGate SNP genotyping analysis. The x-axis represents the chromosomes and the y-axis is the LOD score. Peaks with a LOD score of 3 or higher are considered significant.

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Additional file 2:

Flow cytometric profiles of the MommeDs. (a) Representative flow cytometry profiles show the percentage of GFP-expressing erythrocytes from wild-type and heterozygous mutant littermates (n = 3). The x-axis represents the erythrocyte fluorescence on a logarithmic scale and the y-axis is the number of cells detected at each fluorescence level. (b) Percentage of GFP-expressing cells in wild-type, heterozygous mutant and homozygous mutant (where viable) mice at three weeks of age (mean ± standard error of the mean).

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Additional file 3:

Mapping interval for MommeD30. (a)MommeD30 was produced in the FVB/NJ strain of mice and mapped by crossing twice onto Line3C in a C57BL/6J background. The results of the genotype for seven SNP markers and one microsatellite marker surrounding the linked interval are shown. The number of mice classified into each haplotype is shown on top. Our estimate of the linked interval is between rs29539305 and rs33446195 on chromosome 17 (highlighted). (b) List of genes in the MommeD30 linked interval on chromosome 17. The Mus musculus Ensembl database (release 37) was used to export a list of transcripts (protein coding and non-coding RNAs) within the 1.9 Mbp MommeD30 interval.

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Additional file 4:

GFP expression in offspring of a Rif1GTheterozygote crossed to Line3C. A Rif1 gene trap allele (Rif1GT) had a similar effect on transgene expression as that observed with the MommeD18 mutation, increasing the percentage of expressing cells in mice heterozygous for the gene-trap allele.

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Additional file 5:

Novel mutant alleles of Baz1b, Smchd1, Trim28, Dnmt1 and Smarca5. (a)MommeD16 carries a point mutation resulting in a non-conservative amino acid change in the Wstf domain of Baz1b. (b)MommeD23 and MommeD36 carry point mutations in Smchd1. Both mutations introduce premature stop codons in the Smchd1 protein. (c)MommeD31 carries a point mutation in Trim28 that results in an amino acid change in a highly conserved zinc finger domain. (d)MommeD32 carries a point mutation that results in an amino acid change in the BAH domain of Dnmt1. (e)MommeD35 and MommeD37 carry mutations that result in amino acid changes in highly conserved domains of the Smarca5 protein.

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Additional file 6:

Embryonic development in MommeD mutants. Tabulated data shows the number of observed mice and in brackets the percentage of total. (a) Intercrosses of Pbrm1MommeD27 mice produce no homozygous offspring at three weeks. (b)Brd1MommeD42 homozygous mice are embryonic lethal around 10.5 dpc. (c) Mice carrying mutations in Suv39h1MommeD33 are viable at three weeks. (d)Uhrf1MommeD40 homozygotes are embryonic lethal around 10.5 dpc. (e) Some homozygous Baz1bMommeD16 mice were obtained at three weeks but less than expected. (f) No (Smchd1MommeD36) or few (Smchd1MommeD23) homozygous individuals were recovered from intercrosses. The Smchd1MommeD23 homozygotes that survived were males. (g)Trim28MommeD31 homozygous mice are embryonic lethal prior to 10.5 dpc. (h) Mice homozygous for the Dnmt1MommeD32 mutation die around 10.5 dpc. (i) Intercrosses of Smarca5MommeD35 and Smarca5MommeD37 produced no homozygous offspring at three weeks. Timed matings show that survival of homozygous mutants at 12.5 to 14.5 dpc was rare in Smarca5MommeD35 mutants. In Smarca5MommeD37 mutants homozygous embryonic death occurred prior to 14.5 dpc.

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Additional file 7:

Linkage analysis using Illumina GoldenGate genotyping assay.

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Additional file 8:

List of primer sequences.

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Additional file 9:

Sanger traces.

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