Chem-seq
Identify Sites Bound by Small Chemical Molecules
Chem-seq can be used to detect the binding of small-molecule ligands, such as therapeutic drugs, to proteins associated with the genome. This information may provide important insights into the perturbation of cellular function by small-molecule drugs (Anders et al., 2014).
The Chem-seq method uses 2 approaches. In living cells, the biotinylated drug is added to allow drug-target binding. The complex is crosslinked with formaldehyde, the cells are lysed and sonicated, and the complex is captured on streptavidin beads. The enriched DNA fragments are purified and sequenced. For in vitro analysis, the biotinylated drug is added to a cell extract, and the remaining steps are performed as for the in vivo procedure.
Advantages:
- Can be applied to living, human cells
Disadvantages:
- Creating biotin derivative may alter drug activity
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
None available yet
References:
Anders L., Guenther M. G., Qi J., et al. Genome-wide localization of small molecules. Nat Biotechnol. 2014;32:92-96
Jin C., Yang L., Xie M., et al. Chem-seq permits identification of genomic targets of drugs against androgen receptor regulation selected by functional phenotypic screens. Proc Natl Acad Sci U S A. 2014;111:9235-9240
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History: Chem-seq
Revision by sbrumpton on 2017-06-21 09:28:29 - Show/Hide
Identify Sites Bound by Small Chemical Molecules
Chem-seq can be used to detect the binding of small-molecule ligands, such as therapeutic drugs, to proteins associated with the genome. This information may provide important insights into the perturbation of cellular function by small-molecule drugs (Anders et al., 2014).
The Chem-seq method uses 2 approaches. In living cells, the biotinylated drug is added to allow drug-target binding. The complex is crosslinked with formaldehyde, the cells are lysed and sonicated, and the complex is captured on streptavidin beads. The enriched DNA fragments are purified and sequenced. For in vitro analysis, the biotinylated drug is added to a cell extract, and the remaining steps are performed as for the in vivo procedure.
Advantages:- Can be applied to living, human cells
Disadvantages:- Creating biotin derivative may alter drug activity
Reagents:Illumina Library prep and Array Kit SelectorReviews:None available yet
References:Anders L., Guenther M. G., Qi J., et al. Genome-wide localization of small molecules. Nat Biotechnol. 2014;32:92-96Jin C., Yang L., Xie M., et al. Chem-seq permits identification of genomic targets of drugs against androgen receptor regulation selected by functional phenotypic screens. Proc Natl Acad Sci U S A. 2014;111:9235-9240