ChIP-Chip Designs to Interrogate the Genome of Xenopus Embryos for Transcription Factor Binding and Epigenetic Regulation

被引:6
|
作者
Akkers, Robert C. [1 ]
van Heeringen, Simon J. [1 ]
Manak, J. Robert [2 ,3 ]
Green, Roland D. [2 ]
Stunnenberg, Hendrik G. [1 ]
Veenstra, Gert Jan C. [1 ]
机构
[1] Radboud Univ Nijmegen, Dept Mol Biol, Fac Sci, NL-6525 ED Nijmegen, Netherlands
[2] Roche Nimblegen Inc, Madison, WI USA
[3] Univ Iowa, Dept Biol, Iowa City, IA 52242 USA
来源
PLOS ONE | 2010年 / 5卷 / 01期
基金
美国国家卫生研究院;
关键词
GENE-REGULATION; STEM-CELLS; HISTONE MODIFICATIONS; CHROMATIN STATE; HIGH-RESOLUTION; MAPS; TBP; METHYLATIONS; ROLES; DNA;
D O I
10.1371/journal.pone.0008820
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: 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. Principal Findings: 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. Conclusions: 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.
引用
收藏
页数:7
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