Roof Plate Specific Epigenome Analysis After Maternal Folate Supplementation

该项目旨在研究叶酸对神经管缺陷发展的保护作用是通过表观遗传修饰的假设。对于全基因组CHIP-seq分析，我们首先尝试将标签序列添加到H2 A。Z在体内以细胞类型特异性的方式。然而，C-tetrahydrofuran标记H2 A。Z不组装成染色质。此外，结果表明，在生物素连接酶的过表达中存在细胞毒性，尽管N-末端生物素化H2 A。Z定位于染色质。接下来，我们尝试将标签序列添加到H3.3A，另一种位于转录活性位点的组蛋白变体。包括12个氨基酸接头的C-端羟基磷灰石-或Flag-标记的H3.3源自位于活性染色质部分的34个核苷酸的loxP。我们得出结论，表达C-末端标记的H3.3的载体可用于体内全基因组CHIP-seq分析。

This project was proposed to examine the hypothesis that a protective effect of folate against the development of neural tube defects was via epigenetic modifications. For genome-wide CHIP-seq analysis, we first tried to add tag-sequence to H2A. Z in vivo in a cell-type specific manner. However, C-teminal tagged H2A. Z doesn't assemble to chromatin. In addition, it turned out that there was cellular toxicity in the overexpression of biotin ligase, although N-terminal biotinylated H2A. Z localized at chromatin. We next tried to add tag-sequence to H3.3A, another histone variant localized at transcriptionally active sites. C-teminal HA-or Flag-tagged H3.3 including the 12 amino acids linker originated 34 nt of loxP localized at active chromatin portions. We concluded that the vector expressing the C-terminal tagged H3.3 was useful for genome-wide CHIP-seq analysis in vivo.