目前基因组信息的解码重点已经从基因转向基因调控，即确定参与控制基因适时适地适量表达的调控序列及其所结合的转录因子。ChIP-seq技术虽可鉴定某一转录因子在全基因中的结合靶点，但无法鉴定调控某一兴趣基因的全部转录因子及其所结合的调控序列，而后者对于解析重要功能基因如疾病相关基因的调控机理更为重要。本研究利用CRISPR技术发展一种基因调控元件及因子鉴定新技术，用于高效解码调控某一兴趣基因的全部转录因子及其所结合的调控元件。当研究某一兴趣基因时，设计靶向该基因启动子的一种捕获sgRNA（csgRNA），用dCas9及csgRNA表达载体转染细胞；经培养的细胞用甲醛交联固定，裂解获得细胞核；用转座酶Tn5剪切染色质（tagmentation）；之后用固定寡核苷酸的磁珠捕获Protein-DNA-dCas9/csgRNA复合物，用高通量测序（NGS）及质谱（MS）分析所捕获的DNA及蛋白质。

At present, the focus of decoding of genomic information has shifted from gene to gene regulation, i.e., to determine the regulatory elements (cis elements) harbored in the non-coding sequences occupying 98% of the entire genome and their interacting transcription factors (TFs) (trans factors). These regulatory elements and TFs determine the gene expression at the right place and time at the right dosage. At present, ChIP-seq technology can identify the binding targets of a certain transcription factor in the whole genome, but it cannot identify all transcription factors and their bound regulatory elements that regulate a certain gene of interest, while this is critical for decoding the underlying molecular mechanism controlling important functional genes such as disease-related genes. This study utilizes the rapidly developing CRISPR technology in recent years to develop a novel technology for efficiently identifying all regulatory elements and TFs involved in regulation of an interested gene. This technology has developed a capture-guide RNA (csgRNA) that extends a short sequence at the 3' end of a conventional sgRNA that can anneal to a complementary oligonucleotide immobilized on the surface of the magnetic bead to capture the Protein-DNA-dCas9/sgRNA complex formed in cells. When identifying all regulatory elements and TFs involved in regulation of an interested gene, a csgRNA targeting the promoter of the gene is designed, and dCas9 and csgRNA expression vectors are transfected into cells. The cultured cells are fixed by formaldehyde cross-linking and cytoplasm is removed by lysis. The nuclei is collected and treated with transposase Tn5 for chromatin tagmentation. Subsequently, The Protein-DNA-dCas9/sgRNA complexes are captured with the oligonucleotide-coupled magnetic beads. The DNA and protein in the captured Protein-DNA-dCas9/sgRNA complexes are then analyzed with high-throughput sequencing (NGS) and mass spectrometry (MS) respectively, for determining the entire transcription factors and their bound regulatory elements that regulates a gene of interest.