A Scalable Epitope Tagging Approach for High Throughput ChIP-Seq Analysis

作者：Xiong Xiong; Zhang Yanxiao; Yan Jian; Jain Surbhi; Chee Sora; Ren Bing^*; Zhao Huimin^*

来源：ACS Synthetic Biology, 2017, 6(6): 1034-1042.

DOI：10.1021/acssynbio.6b00358

• ChIP-Seq
• microhomology mediated end joining
• CRISPR/Cas9
• genome engineering
• FLAG tagging

Eukaryotic transcriptional factors (TFs) typically recognize short genomic sequences alone or together with other proteins to modulate gene expression. Mapping of TF-DNA interactions in the genome is crucial for understanding the gene regulatory programs in cells. While chromatin immunoprecipitation followed by sequencing (ChIP-Seq) is commonly used for this purpose, its application is severely limited by the availability of suitable antibodies for TFs. To overcome this limitation, we developed an efficient and scalable strategy named cmChIP-Seq that combines the clustered regularly interspaced short palindromic repeats (CRISPR) technology with microhomology mediated end joining (MMEJ) to genetically engineer a TF with an epitope tag. We demonstrated the utility of this tool by applying it to four TFs in a human colorectal cancer cell line. The highly scalable procedure makes this strategy ideal for ChIP-Seq analysis of TFs in diverse species and cell types.

• 出版日期2017-6