Chemogenomic profiling is a powerful and unbiased approach to elucidate pharmacological targets and the mechanism of bioactive compounds. It is based on identifying cellular hypersensitivity and resistance caused by individual gene modulations with genome-wide coverage. Due to the requirement of bar-coded, genome-wide deletion collections, high-resolution experiments of this nature have historically been limited to fungal systems. Pooled RNAi reagents have enabled similar attempts in mammalian cells but efforts have been hampered by significant off-target effects and experimental noise. The CRISPR/Cas9 system for the first time enables precise DNA editing at defined loci in a genome-wide fashion. Here we present the detailed protocol that leverages the CRISPR/Cas9 system for chemogenomic profiling and target identification of diverse chemical probes.
CITATION STYLE
Hoepfner, D., McAllister, G., & Hoffman, G. R. (2019). CRISPR/Cas9-based chemogenomic profiling in mammalian cells. In Methods in Molecular Biology (Vol. 1888, pp. 153–174). Humana Press Inc. https://doi.org/10.1007/978-1-4939-8891-4_9
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