DNA Methylation Assay for X-Chromosome Inactivation in Female Human iPS Cells
DNA Methylation Assay for X-Chromosome Inactivation in Female Human iPS Cells
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DOI：
10.1007/s12015-011-9238-6
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发表时间：
2011-11-01
影响因子：
4.8
通讯作者：
Mongomery, Karen Dyer
中科院分区：
文献类型：
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作者：
Kiedrowski, Lesli A.;Raca, Gordana;Mongomery, Karen Dyer
对人类诱导多能干细胞（iPS）的表观遗传状态的极大兴趣激发了对其X失活模式的大量研究。然而，迄今为止，在未分化的iPS细胞中已经描述了X失活的存在和不存在。不同研究之间结果不一致的原因尚不清楚，需要对所有雌性人iPS细胞系进行进一步的X-灭活试验。目前数据中的一些不一致很可能是由于不同作者使用不同的X-灭活测定法造成的。我们提供了一个详细的协议，一个简单的，可靠的和负担得起的X-失活分析的基础上启动子甲基化和CAG重复多态性的人雄激素受体（AR）基因在Xq11.2。该检测方法通常用于临床遗传实验室，我们建议它可能是理想的常规评估和监测女性人iPS细胞系中的X-失活状态。
Remarkable interest in the epigenetic status of human induced pluripotent stem (iPS) cells inspired numerous studies of their X-inactivation patterns. However, both the presence and the absence of X-inactivation have been described to date in undifferentiated iPS cells. The reasons for the discordant results between different studies are unclear, and further X-inactivation testing is warranted for all female human iPS cell lines. Some of the inconsistency in the current data most likely results from the use of different X-inactivation assays by different authors. We provide a detailed protocol for a simple, reliable and affordable X-inactivation assay based on promoter methylation and CAG-repeat polymorphism in the human androgen receptor (AR) gene at Xq11.2. This assay is commonly used in clinical genetic laboratories and we propose that it could be ideal for routine assessment and monitoring of the X-inactivation status in female human iPS cell lines.