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Targeted mutation of the DNA methyltransferase gene results in embryonic lethality
Abstract
Gene targeting in embryonic stem (ES) cells has been used to mutate the murine DNA methyltransferase gene. ES cell lines homozygous for the mutation were generated by consecutive targeting of both wild-type alleles; the mutant cells were viable and showed no obvious abnormalities with respect to growth rate or morphology, and had only trace levels of DNA methyltransferase activity. A quantitative end-labeling assay showed that the level of m5C in the DNA of homozygous mutant cells was about one-third that of wild-type cells, and Southern blot analysis after cleavage of the DNA with a methylation-sensitive restriction endonuclease revealed substantial demethylation of endogenous retroviral DNA. The mutation was introduced into the germline of mice and found to cause a recessive lethal phenotype. Homozygous embryos were stunted, delayed in development, and did not survive past mid-gestation. The DNA of homozygous embryos showed a reduction of the level of m5C similar to that of homozygous ES cells. These results indicate that while a 3-fold reduction in levels of genomic m5C has no detectable effect on the viability or proliferation of ES cells in culture, a similar reduction of DNA methylation in embryos causes abnormal development and embryonic lethality.
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“Global deletion of mouse Dnmt1, Dnmt3b, or both Dnmt3a + Dnmt3b results in midgestation lethality [gestational day (GD) 9.5–11.5], while deletion of only Dnmt3a produces severe growth retardation and lethality by 4 weeks of age [ 70–72 ].”
From DNA methylation: a mechanism linking environmental chemical exposures to risk of autism spectrum disorders? (2016) · cites this paper as [70]
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Metadata
- Authors
- Li, E., Bestor, T. H., & Jaenisch, R.
- Affiliations
- Whitehead Institute for Biomedical Research (US)
- Harvard University (US)
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 1992-06
- Volume
- 69
- Issue
- 6
- Pages
- 915-926
- ISSN
- 0092-8674 (ISSN-L)
- OpenAlex
- W2046241907
- Cited by
- 3,984
- References
- 61
- Citations by year
- 2026: 50 · 2025: 60 · 2024: 81 · 2023: 84 · 2022: 84 · 2021: 90 · 2020: 95 · 2019: 127 · 2018: 137 · 2017: 113 · 2016: 147 · 2015: 157 · 2014: 169 · 2013: 178 · 2012: 187
- FWCI
- 28.04
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Cell (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r35ca044339-13, 5r01gm043565-04)
- National Cancer Institute (R35 CA 44339-05)
- National Institute of General Medical Sciences (GM-43565)
- 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 › Genetics › Animal Genetics and Reproduction
- Keywords
- DNA methyltransferases, embryonic lethality, DNA methylation, 5-methylcytosine, demethylation, embryonic stem cells, germline mutations, recessive lethality, mouse embryo development, gene targeting
- MeSH terms
- Animals, Chimera, DNA, DNA (Cytosine-5-)-Methyltransferases, Genes, Lethal, Homozygote, Methylation, Stem Cells, Blotting, Western, Mutagenesis, Insertional, Mice
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