NET-Seq

Chem-Seq

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. 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.

Pros:

• Can be applied to living, human cells

Cons:

• Creating biotin derivative may alter drug activity

• Anders L., Guenther M. G., Qi J., et al. Genome-wide localization of small molecules. Nat Biotechnol. 2014;32:92-96

1. Anders L., Guenther M. G., Qi J., et al. Genome-wide localization of small molecules. Nat Biotechnol. 2014;32:92-96
2. 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