Chamberlain Stormy J, Germain Noelle D, Chen Pin-Fang, Hsiao Jack S, Glatt-Deeley Heather
University of Connecticut Health Center, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA.
Methods Mol Biol. 2016;1353:45-64. doi: 10.1007/7651_2014_169.
Induced pluripotent stem cell (iPSC) technology has allowed for the invaluable modeling of many genetic disorders including disorders associated with genomic imprinting. Genomic imprinting involves differential DNA and histone methylation and results in allele-specific gene expression. Most of the epigenetic marks in somatic cells are erased and reestablished during the process of reprogramming into iPSCs. Therefore, in generating models of disorders associated with genomic imprinting, it is important to verify that the imprinting status and allele-specific gene expression patterns of the parental somatic cells are maintained in their derivative iPSCs. Here, we describe three techniques: DNA methylation analysis, allele-specific PCR, and RNA FISH, which we use to analyze genomic imprinting in iPSC models of neurogenetic disorders involving copy number variations of the chromosome 15q11-q13 region.
诱导多能干细胞（iPSC）技术使得对许多遗传疾病进行极为重要的建模成为可能，这些疾病包括与基因组印记相关的疾病。基因组印记涉及DNA和组蛋白的差异甲基化，并导致等位基因特异性基因表达。在重编程为iPSC的过程中，体细胞中的大多数表观遗传标记会被擦除并重新建立。因此，在生成与基因组印记相关的疾病模型时，重要的是要验证亲代体细胞的印记状态和等位基因特异性基因表达模式在其衍生的iPSC中是否得以维持。在此，我们描述了三种技术：DNA甲基化分析、等位基因特异性PCR和RNA荧光原位杂交，我们使用这些技术来分析涉及15q11 - q13染色体区域拷贝数变异的神经遗传疾病的iPSC模型中的基因组印记。
