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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Sensitive ChIP-DSL technology reveals an extensive estrogen receptor α-binding program on human gene promoters
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Sensitive ChIP-DSL technology reveals an extensive estrogen receptor α-binding program on human gene promoters

机译：敏感的ChIP-DSL技术揭示了人类基因启动子上广泛的雌激素受体α结合程序

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ChIP coupled with microarray provides a powerful tool to determine in vivo binding profiling of transcription factors to deduce regulatory circuitries in mammalian cells. Aiming at improving the specificity and sensitivity of such analysis, we developed a new technology called ChIP-DSL using the DNA selection and ligation (DSL) strategy, permitting robust analysis with much reduced materials compared with standard procedures. We profiled general and sequence-specific DNA binding transcription factors using a full human genome promoter array based on the ChIP-DSL technology, revealing an unprecedented number of the estrogen receptor (ERα) target genes in MCF-7 cells. Coupled with gene expression profiling, we found that only a fraction of these direct ERa target genes were highly responsive to estrogen and that the expression of those ERα-bound, estrogen-inducible genes was associated with breast cancer progression in humans. This study demonstrates the power of the ChIP-DSL technology in revealing regulatory gene expression programs that have been previously invisible in the human genome.
机译：ChIP与微阵列相结合提供了一种强大的工具，可用于确定转录因子的体内结合谱，以推断哺乳动物细胞中的调节回路。为了提高此类分析的特异性和敏感性，我们使用DNA选择和连接（DSL）策略开发了一种称为ChIP-DSL的新技术，与标准程序相比，该方法可使用大量减少的材料进行可靠的分析。我们使用了基于ChIP-DSL技术的完整人类基因组启动子阵列，分析了一般的和序列特异性的DNA结合转录因子，揭示了MCF-7细胞中前所未有的雌激素受体（ERα）靶基因数量。结合基因表达谱分析，我们发现这些直接的ERa靶基因中只有一小部分对雌激素高度敏感，而那些与ERα结合的雌激素诱导型基因的表达与人类乳腺癌的进展有关。这项研究证明了ChIP-DSL技术在揭示以前在人类基因组中不可见的调节基因表达程序的能力。

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