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An N -ethyl- N -nitrosourea screen for genes involved in variegation in the mouse
Open accessAbstract
We have developed a sensitized screen to identify genes involved in gene silencing, using random N -ethyl- N -nitrosourea mutagenesis on mice carrying a variegating GFP transgene. The dominant screen has produced six mutant lines, including both suppressors and enhancers of variegation. All are semidominant and five of the six are homozygous embryonic lethal. In one case, the homozygous lethality depends on sex: homozygous females die at midgestation and display abnormal DNA methylation of the X chromosome, whereas homozygous males are viable. Linkage analysis reveals that the mutations map to unique chromosomal locations. We have studied the effect of five of the mutations on expression of an endogenous allele known to be sensitive to epigenetic state, agouti viable yellow . In all cases, there is an effect on penetrance, and in most cases, parent of origin and sex-specific effects are detected. This screen has identified genes that are involved in epigenetic reprogramming of the genome, and the behavior of the mutant lines suggests a common mechanism between X inactivation and transgene and retrotransposon silencing. Our findings raise the possibility that the presence or absence of the X chromosome in mammals affects the establishment of the epigenetic state at autosomal loci by acting as a sink for proteins involved in gene silencing. The study demonstrates the power of sensitized screens in the mouse not only for the discovery of novel genes involved in a particular process but also for the elucidation of the biology of that process.
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“A preliminary description of the first six heritable mutations, which we refer to as Modifiers of murine metastable epialleles or Mommes, following the screening of 608 G1 offspring, has been published [10].”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“We reported that all six were dosage-effect genes and five of the six were homozygous lethal, with loss of homozygotes apparent at weaning, but no knowledge of when death occurred.”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“We have previously reported that the GFP transgene used in this screen has integrated as an approximately 11 copy array on chromosome 1 [10].”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“Recently, we established a similar screen in the mouse using a transgenic line that expresses green fluorescent protein (GFP) in a variegated manner in erythrocytes [10].”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“MommeD2 and MommeD4 are homozygous lethal at 8.5 days post-coitus (dpc) and 17.5 dpc, respectively [10,12].”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“We have reported previously that as the percentage of expressing cells increases, the mean fluorescence of the expressing cells increases [10].”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“MommeD5 was localized to a 1.42 Mb region on chromosome 4 flanked by the markers rs27486641 and rs27541967 [10] (Table 4).”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“MommeD1 is homozygous lethal in females only, with death occurring around mid-gestation [10,11].”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“The frequency with which we found these mutations, 1 in 100 G1 offspring, was similar to our previous results [10].”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
In the citing paper
“Extension of the screen has produced four new MommeDs, MommeD7-D10, all of which behave in a semidominant fashion, as do the six previously reported [10].”
From A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development (2008) · cites this paper as [10]
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Zhuang, Q. K.-W., Bauermeister, K., Gálvez, J. H., AlOgayil, N., Batdorj, E., de Villena, F. P., Taketo, T., Bourque, G., & Naumova, A. K. (2024) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2024.09.10.612032
Desai, V. P., Chouaref, J., Wu, H., Pastor, W. A., Kan, R. L., Oey, H., Li, Z., Ho, J., Vonk, K. K. D., León, D. S., et al. (2021) · Figshare · doi:10.6084/m9.figshare.16893220
Professor Emma Whitelaw, a pioneer of epigenetic inheritance
Ashe, A., & Hawes, J. J. (2026) · Frontiers in Epigenetics and Epigenomics · doi:10.3389/freae.2025.1733711
Self-citationBiogenesis and Function of H3K9me3 at telomeres of mouse embryonic stem cells
Kan, S. (2015) · theses.fr (ABES) · doi:10.70675/cbaee4b0z64cez4b4eza882zbaf2c49e74a0
La dystrophie facio-scapulo-humérale : du diagnostic aux aspects fonctionnels
Delourme, M. (2022) · theses.fr (ABES) · doi:10.70675/8a7663bbzddd9z4e58zbf4fza999c4a9a715
Effet de position télomérique : implications dans la régulation du génome et les maladies rares
Chevalier, R. (2022) · theses.fr (ABES) · doi:10.70675/89769afeza443z495cz9933z31ab8621510e
Metadata
- Authors
- Blewitt, M. E., Vickaryous, N. K., Hemley, S. J., Ashe, A., Bruxner, T. J., Preis, J. I., Arkell, R., & Whitelaw, E.
- Affiliations
- The University of Sydney (AU)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2005-05-12
- Volume
- 102
- Issue
- 21
- Pages
- 7629-7634
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC1140414
- OpenAlex
- W2089778660
- Cited by
- 186
- References
- 37
- Citations by year
- 2026: 1 · 2025: 6 · 2024: 9 · 2023: 4 · 2022: 8 · 2021: 11 · 2020: 5 · 2019: 11 · 2018: 16 · 2017: 17 · 2016: 8 · 2015: 9 · 2014: 6 · 2013: 14 · 2012: 11
- FWCI
- 3.39
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- ANU Open Research (Australian National University) (submittedVersion)
- Funders
- Medical Research Council
- National Health and Medical Research Council (301955)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic Syndromes and Imprinting
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- variegation, gene silencing, N-ethyl-N-nitrosourea mutagenesis, GFP transgene, X chromosome inactivation, epigenetic reprogramming, retrotransposon silencing, transposon silencing, embryonic lethality, parent-of-origin effects, penetrance, mouse mutants
- MeSH terms
- Alleles, Animals, Chromosome Mapping, Ethylnitrosourea, Flow Cytometry, Genetic Testing, Mice, Transgenic, Phenotype, Pigmentation, Sex Factors, X Chromosome, Mutagenesis, Transgenes, DNA Methylation, Gene Silencing, Epigenesis, Genetic, Green Fluorescent Proteins, Mice
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