Disruption of imprinting and aberrant embryo development in completely inbred embryonic stem cell-derived mice.
The completely embryonic stem (ES) cell-derived mice (ES mice) produced by tetraploid embryo complementation provide us with a rapid and powerful approach for functional genome analysis. However, inbred ES cell lines often fail to generate ES mice. The genome of mouse ES cells is extremely unstable during in vitro culture and passage, and the expression of the imprinted genes is most likely to be affected. Whether the ES mice retain or repair the abnormalities of the donor ES cells has still to be determined. Here we report that the inbred ES mice were efficiently produced with the inbred ES cell line (SCR012). The ES fetuses grew more slowly before day 17.5 after mating, but had an excessive growth from day 17.5 to birth. Five imprinted genes examined (H19, Igf2, Igf2r, Peg1, Peg3) were expressed abnormally in ES fetuses. Most remarkably, the expression of H19 was dramatically repressed in the ES fetuses through the embryo developmental stage, and this repression was associated with abnormal biallelic methylation of the H19 upstream region. The altered methylation pattern of H19 was further demonstrated to have arisen in the donor ES cells and persisted on in vivo differentiation to the fetal stage. These results indicate that the ES fetuses did retain the epigenetic alterations in imprinted genes from the donor ES cells.
Deng
Tao
T
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology and Model Organism Research Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Kuang
Ying
Y
Zhang
Dan
D
Wang
Long
L
Sun
Ruilin
R
Xu
Guojiang
G
Wang
Zhugang
Z
Fei
Jian
J
D000818:Animals; D001483:Base Sequence; D019175:DNA Methylation; D017931:DNA Primers; D047108:Embryonic Development; D053595:Embryonic Stem Cells / Q000166:cytology*; D018392:Genomic Imprinting; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D010641:Phenotype
D017931:DNA Primers