Kwon Young-Soo, Garcia-Bassets Ivan, Hutt Kasey R, Cheng Christine S, Jin Mingjie, Liu Dongyan, Benner Chris, Wang Dong, Ye Zhen, Bibikova Marina, Fan Jian-Bing, Duan Lingxun, Glass Christopher K, Rosenfeld Michael G, Fu Xiang-Dong
Department of Cellular and Molecular Medicine, University of California at San Diego School of Medicine, La Jolla, CA 92093-0651, USA.
Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):4852-7. doi: 10.1073/pnas.0700715104. Epub 2007 Mar 14.
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 (ERalpha) target genes in MCF-7 cells. Coupled with gene expression profiling, we found that only a fraction of these direct ERalpha target genes were highly responsive to estrogen and that the expression of those ERalpha-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.
染色质免疫沉淀技术(ChIP)与微阵列相结合,为确定转录因子在体内的结合图谱以推断哺乳动物细胞中的调控通路提供了一个强大的工具。为了提高此类分析的特异性和灵敏度,我们利用DNA选择与连接(DSL)策略开发了一种名为ChIP-DSL的新技术,与标准程序相比,该技术可以使用更少的材料进行稳健的分析。我们基于ChIP-DSL技术,使用全人类基因组启动子阵列对通用和序列特异性DNA结合转录因子进行了分析,揭示了MCF-7细胞中数量空前的雌激素受体(ERalpha)靶基因。结合基因表达谱分析,我们发现这些直接的ERalpha靶基因中只有一小部分对雌激素高度敏感,并且那些与ERalpha结合、雌激素诱导的基因的表达与人类乳腺癌进展相关。这项研究证明了ChIP-DSL技术在揭示人类基因组中以前不可见的调控基因表达程序方面的强大作用。