Abstract:
: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.
journal_name
ACS Synth Bioljournal_title
ACS synthetic biologyauthors
Xiong X,Zhang Y,Yan J,Jain S,Chee S,Ren B,Zhao Hdoi
10.1021/acssynbio.6b00358subject
Has Abstractpub_date
2017-06-16 00:00:00pages
1034-1042issue
6issn
2161-5063journal_volume
6pub_type
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