Department of Molecular Biology, Faculty of Science, Radboud University Nijmegen, Nijmegen, The Netherlands.
PLoS One. 2010 Jan 21;5(1):e8820. doi: 10.1371/journal.pone.0008820.
Chromatin immunoprecipitation combined with genome tile path microarrays or deep sequencing can be used to study genome-wide epigenetic profiles and the transcription factor binding repertoire. Although well studied in a variety of cell lines, these genome-wide profiles have so far been little explored in vertebrate embryos.
Here we report on two genome tile path ChIP-chip designs for interrogating the Xenopus tropicalis genome. In particular, a whole-genome microarray design was used to identify active promoters by close proximity to histone H3 lysine 4 trimethylation. A second microarray design features these experimentally derived promoter regions in addition to currently annotated 5' ends of genes. These regions truly represent promoters as shown by binding of TBP, a key transcription initiation factor.
A whole-genome and a promoter tile path microarray design was developed. Both designs can be used to study epigenetic phenomena and transcription factor binding in developing Xenopus embryos.
染色质免疫沉淀结合基因组平铺路径微阵列或深度测序可用于研究全基因组表观遗传谱和转录因子结合谱。尽管在多种细胞系中得到了很好的研究,但这些全基因组图谱在脊椎动物胚胎中迄今尚未得到充分探索。
本文报道了两种用于研究非洲爪蟾(Xenopus tropicalis)基因组的基因组平铺路径 ChIP-chip 设计。特别是,全基因组微阵列设计用于通过与组蛋白 H3 赖氨酸 4 三甲基化的接近程度来鉴定活性启动子。第二个微阵列设计除了当前注释的基因 5' 端外,还具有这些经过实验衍生的启动子区域。这些区域如关键转录起始因子 TBP 的结合所示,确实代表启动子。
开发了全基因组和启动子平铺路径微阵列设计。这两种设计都可用于研究发育中的非洲爪蟾胚胎中的表观遗传现象和转录因子结合。