In vivo characterization of regulatory polymorphisms by allele-specific quantification of RNA polymerase loading
In vivo characterization of regulatory polymorphisms by allele-specific quantification of RNA polymerase loading
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DOI：
10.1038/ng1124
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发表时间：
2003-04-01
期刊：
影响因子：
30.8
通讯作者：
Kwiatkowski, DR
中科院分区：
文献类型：
--
作者：
Knight, JC;Keating, BJ;Kwiatkowski, DR
调控多态性的体内表征是下一代人类遗传分析的关键要求。在这里，我们描述haploChIP，一种方法，使用染色质免疫沉淀（ChIP）和质谱鉴定差异蛋白质-DNA结合在体内与等位基因变异的基因。我们证明了这种方法与印迹基因SNRPN。HaploChIP显示SNRPN位点结合的磷酸化RNA聚合酶II水平与等位基因特异性表达密切相关。该方法应用于TNF/LTA位点，确定了与LTA等位基因特异性转录相关的功能重要的单倍型。haploChIP方法可用于高通量筛选影响体内基因调控的常见DNA多态性。
In vivo characterization of regulatory polymorphisms is a key requirement for next-generation human genetic analysis. Here we describe haploChIP, a method that uses chromatin immunoprecipitation (ChIP) and mass spectrometry to identify differential protein-DNA binding in vivo associated with allelic variants of a gene. We demonstrate this approach with the imprinted gene SNRPN. HaploChIP showed close correlation between the level of bound phosphorylated RNA polymerase II at the SNRPN locus and allele-specific expression. Application of the approach to the TNF/LTA locus identified functionally important haplotypes that correlate with allele-specific transcription of LTA. The haploChIP method may be useful in high-throughput screening for common DNA polymorphisms that affect gene regulation in vivo.