Disruption of imprinting and aberrant embryo development in completely inbred embryonic stem cell-derived mice
Disruption of imprinting and aberrant embryo development in completely inbred embryonic stem cell-derived mice
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DOI：
10.1111/j.1440-169x.2007.00955.x
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发表时间：
2007-09-01
影响因子：
2.5
通讯作者：
Fei, Jian
中科院分区：
文献类型：
--
作者：
Deng, Tao;Kuang, Ying;Fei, Jian
通过四倍体胚胎互补产生的全胚胎干细胞来源小鼠（ES小鼠）为功能基因组分析提供了一种快速而有力的方法。然而，近交的胚胎干细胞系经常不能产生胚胎干细胞小鼠。小鼠胚胎干细胞的基因组在体外培养和传代过程中非常不稳定，印迹基因的表达最容易受到影响。胚胎干细胞小鼠是否保留或修复供体胚胎干细胞的异常还有待确定。本文报道了用近交系胚胎干细胞（SCR012）高效培育近交系胚胎干细胞小鼠。胚胎在交配后17.5天前生长较慢，但在17.5天至出生时生长过快。5个印迹基因（H19、Igf2、Igf2r、Peg1、Peg3）在ES胎儿中表达异常。最值得注意的是，在胚胎发育阶段，H19的表达在ES胚胎中被显著抑制，这种抑制与H19上游区域的双等位基因甲基化异常有关。H19甲基化模式的改变在供体胚胎干细胞中出现，并持续在体内分化至胎儿期。这些结果表明胚胎干细胞胚胎确实保留了来自供体胚胎干细胞的印迹基因的表观遗传改变。
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.