A reporter model to visualize imprinting stability at the Dlk1 locus during mouse development and in pluripotent cells.
A reporter model to visualize imprinting stability at the Dlk1 locus during mouse development and in pluripotent cells.
复制标题
DOI：
10.1242/dev.138255
复制
发表时间：
2016-11-15
期刊：
影响因子：
--
通讯作者：
Stadtfeld M
中科院分区：
文献类型：
--
作者：
Swanzey E;Stadtfeld M
基因组印记导致编码生长和增殖的重要调节因子的基因的单等位基因表达。印迹基因（如Dlk 1-Dio 3基因座内的印迹基因）的失调与发育综合征和特定疾病相关。由于缺乏合适的模型系统，我们询问印迹不稳定性原因的能力受到阻碍。在这里，我们描述了Dlk 1基因敲入报告小鼠，使单细胞可视化等位基因特异性表达和前瞻性的细胞分离，同时。我们表明，这种“印迹报告小鼠”可用于检测发育中胚胎的组织特异性Dlk 1表达模式。我们还将该系统应用于多能细胞培养，并证明它忠实地表明细胞重编程后诱导的DNA甲基化变化。最后，报告系统揭示了在长期培养和体外分化过程中，氧水平升高在侵蚀印迹Dlk 1表达中的作用。研究等位基因特异性表达的可能性，在不同的情况下，使我们的报告系统的一个有用的工具，解剖基因组印记在正常发育和疾病的调节。总结：Dlk 1基因敲入报告小鼠报告等位基因和组织特异性Dlk 1表达在发育中的胚胎，可用于研究细胞重编程过程中基因组印记的变化。
Genomic imprinting results in the monoallelic expression of genes that encode important regulators of growth and proliferation. Dysregulation of imprinted genes, such as those within the Dlk1-Dio3 locus, is associated with developmental syndromes and specific diseases. Our ability to interrogate causes of imprinting instability has been hindered by the absence of suitable model systems. Here, we describe a Dlk1 knock-in reporter mouse that enables single-cell visualization of allele-specific expression and prospective isolation of cells, simultaneously. We show that this ‘imprinting reporter mouse’ can be used to detect tissue-specific Dlk1 expression patterns in developing embryos. We also apply this system to pluripotent cell culture and demonstrate that it faithfully indicates DNA methylation changes induced upon cellular reprogramming. Finally, the reporter system reveals the role of elevated oxygen levels in eroding imprinted Dlk1 expression during prolonged culture and in vitro differentiation. The possibility to study allele-specific expression in different contexts makes our reporter system a useful tool to dissect the regulation of genomic imprinting in normal development and disease. Summary: A Dlk1 knock-in reporter mouse reports allele- and tissue-specific Dlk1 expression in developing embryos that can be used to study changes in genomic imprinting during cellular reprogramming.
登录
查看更多内容
影响因子：
1.8
作者：
Wu, Qiong;Kawahara, Manabu;Kono, Tomohiro
通讯作者：
Kono, Tomohiro
影响因子：
4.8
作者：
Weng, Tingting;Gao, Li;Liu, Lin
通讯作者：
Liu, Lin
影响因子：
10.5
作者：
Ginart P;Kalish JM;Jiang CL;Yu AC;Bartolomei MS;Raj A
通讯作者：
Raj A
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
64.8
作者：
通讯作者：
--