Gene Expression and Methylation Patterns in Cloned Embryos.
From: Department of Biotechnology, Institute for Animal Science, FAL, Mariensee, Neustadt, Germany.
Methods in molecular biology (Clifton, N.J.)
- Publish Date: 2006
- ISSN: 1064-3745
- Volume: 348
- Issue:
- Pages: 285-304
- Medium: Print
- Language: English
- Citation (JAMA): Wrenzycki Christine, Herrmann Doris, Gebert Claudia, et al. Gene Expression and Methylation Patterns in Cloned Embryos.. Methods Mol. Biol. 2006;348:285-304
Abstract
A considerable proportion of the offspring, in particular in ruminants and mouse, born from nuclear transfer (NT)-derived and in vitro-produced (IVP) embryos are affected by multiple abnormalities, of which a high birthweight and an extended gestation length are the predominant features; a phenomenon that has been termed “Large Offspring syndrome” (LOS). According to a current hypothesis, LOS is caused by persistent aberrations of expression patterns of developmentally important genes starting as early as at the preimplantation stages. The underlying mechanisms are widely unknown at present, but epigenetic modifications of embryonic and fetal gene expression patterns, primarily caused by alterations in DNA methylation are thought to be involved in this syndrome. Appropriate DNA methylation is essential for regular transcription during mammalian development and differentiation. Sensitive reverse transcription polymerase chain reaction assays allow the study of messenger RNA (mRNA) expression levels of specific genes in single embryos. The methylation status of a specific gene can be assessed by bisulfite sequencing. Studies to unravel mRNA expression patterns from IVP- and NT-derived embryos have revealed numerous aberrations ranging from suppression of expression to de novo overexpression or more frequently to a significant upregulation or downregulation of a specific gene. mRNA expression patterns from in vivo-derived embryos are essential as the “physiological standard” against which the findings for IVP and NT-derived embryos are to be compared. Unraveling the underlying molecular mechanisms will contribute to the production of viable embryos and aid to improve biotechnologies applied to early mammalian embryos.
Mesh Headings (Keywords): Animals, Cell Nucleus, Cloning, Organism, DNA Methylation, Embryo, Mammalian, Epigenesis, Genetic, Fertilization in Vitro, Gene Expression, Mice, RNA, Messenger, Reverse Transcriptase Polymerase Chain Reaction, Sequence Analysis, DNA
Check for Full Text / PubMed Unique Identifier (PMID): 16988388
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