2B-08 Analysis of DNA methylation for establishing of imprinted expression of Igf2r gene using in vitro differentiation system of mouse embryonic stem (ES) cells

Generate imprinting is essential for normal development of mammalian embryos. Parent of origin specific expression of imprinted genes is established at the certain period of early embryonic development. We are using newly established mouse ES cell lines derived from inter-subspecies F1 hybrid mouse...

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Published in:Genes & Genetic Systems Vol. 85; no. 6; p. 407
Main Authors: MISE Nathan, KONDO Masayo, ABE Kuniya
Format: Journal Article
Language:Japanese
Published: The Genetics Society of Japan 2010
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Abstract Generate imprinting is essential for normal development of mammalian embryos. Parent of origin specific expression of imprinted genes is established at the certain period of early embryonic development. We are using newly established mouse ES cell lines derived from inter-subspecies F1 hybrid mouse embryos between C57BL/6J and MSM/Ms those allow us to distinguish parental origin of genome and transcripts using abundant SNPs existing between these two inbred lines to analyze detailed molecular mechanisms of imprinted expression pattern. Although the differential methylation pattern of Igf2r differentially methylated region (DMR) is maintained in ES cells, Igf2r mRNA was expressed from both alleles in undifferentiated ES cells. The maternal expression was established accompanying in vitro differentiation of ES cells. The secondary DMR of Igf2r gene that locates in its promoter region was free of methylation in undifferentiated state ES cells, but methylation of the promoter occurred following cell differentiation. These results indicate that our in vitro differentiating system of ES cells reproduce in vivo events and suit to study establishing imprinted expression pattern in vitro.
AbstractList Generate imprinting is essential for normal development of mammalian embryos. Parent of origin specific expression of imprinted genes is established at the certain period of early embryonic development. We are using newly established mouse ES cell lines derived from inter-subspecies F1 hybrid mouse embryos between C57BL/6J and MSM/Ms those allow us to distinguish parental origin of genome and transcripts using abundant SNPs existing between these two inbred lines to analyze detailed molecular mechanisms of imprinted expression pattern. Although the differential methylation pattern of Igf2r differentially methylated region (DMR) is maintained in ES cells, Igf2r mRNA was expressed from both alleles in undifferentiated ES cells. The maternal expression was established accompanying in vitro differentiation of ES cells. The secondary DMR of Igf2r gene that locates in its promoter region was free of methylation in undifferentiated state ES cells, but methylation of the promoter occurred following cell differentiation. These results indicate that our in vitro differentiating system of ES cells reproduce in vivo events and suit to study establishing imprinted expression pattern in vitro.
Author KONDO Masayo
ABE Kuniya
MISE Nathan
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CorporateAuthor Dept. Pharmacology
Sch. Med
RIKEN
Div. Env. Toxicology
BioResource Centre
Jichi Medical Univ
Mammalian Cellular Dynamics
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Title 2B-08 Analysis of DNA methylation for establishing of imprinted expression of Igf2r gene using in vitro differentiation system of mouse embryonic stem (ES) cells
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