
Target identification of small molecules using large-scale CRISPR-Cas mutagenesis scanning of essential genes
- Author
- Jasper Edgar Neggers, Bert Kwanten, Tim Dierckx, Hiroki Noguchi, Arnout Voet, Lotte Bral, Kristien Minner, Bob Massant, Nicolas Kint (UGent) , Michel Delforge, Thomas Vercruysse, Erkan Baloglu, William Senapedis, Maarten Jacquemyn and Dirk Daelemans
- Organization
- Abstract
- Unraveling the mechanism of action and molecular target of small molecules remains a major challenge in drug discovery. While many cancer drugs target genetic vulnerabilities, loss-of-function screens fail to identify essential genes in drug mechanism of action. Here, we report CRISPRres, a CRISPR-Cas-based genetic screening approach to rapidly derive and identify drug resistance mutations in essential genes. It exploits the local genetic variation created by CRISPR-Cas-induced non-homologous end-joining (NHEJ) repair to generate a wide variety of functional in-frame mutations. Using large sgRNA tiling libraries and known drug-target pairs, we validate it as a target identification approach. We apply CRISPRres to the anticancer agent KPT-9274 and identify nicotinamide phosphoribosyltransferase (NAMPT) as its main target. These results present a powerful and simple genetic approach to create many protein variants that, in combination with positive selection, can be applied to reveal the cellular target of small-molecule inhibitors.
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-01H1Y107JN3JTNKNZRZW6TZXCB
- MLA
- Neggers, Jasper Edgar, et al. “Target Identification of Small Molecules Using Large-Scale CRISPR-Cas Mutagenesis Scanning of Essential Genes.” NATURE COMMUNICATIONS, vol. 9, 2018, doi:10.1038/s41467-017-02349-8.
- APA
- Neggers, J. E., Kwanten, B., Dierckx, T., Noguchi, H., Voet, A., Bral, L., … Daelemans, D. (2018). Target identification of small molecules using large-scale CRISPR-Cas mutagenesis scanning of essential genes. NATURE COMMUNICATIONS, 9. https://doi.org/10.1038/s41467-017-02349-8
- Chicago author-date
- Neggers, Jasper Edgar, Bert Kwanten, Tim Dierckx, Hiroki Noguchi, Arnout Voet, Lotte Bral, Kristien Minner, et al. 2018. “Target Identification of Small Molecules Using Large-Scale CRISPR-Cas Mutagenesis Scanning of Essential Genes.” NATURE COMMUNICATIONS 9. https://doi.org/10.1038/s41467-017-02349-8.
- Chicago author-date (all authors)
- Neggers, Jasper Edgar, Bert Kwanten, Tim Dierckx, Hiroki Noguchi, Arnout Voet, Lotte Bral, Kristien Minner, Bob Massant, Nicolas Kint, Michel Delforge, Thomas Vercruysse, Erkan Baloglu, William Senapedis, Maarten Jacquemyn, and Dirk Daelemans. 2018. “Target Identification of Small Molecules Using Large-Scale CRISPR-Cas Mutagenesis Scanning of Essential Genes.” NATURE COMMUNICATIONS 9. doi:10.1038/s41467-017-02349-8.
- Vancouver
- 1.Neggers JE, Kwanten B, Dierckx T, Noguchi H, Voet A, Bral L, et al. Target identification of small molecules using large-scale CRISPR-Cas mutagenesis scanning of essential genes. NATURE COMMUNICATIONS. 2018;9.
- IEEE
- [1]J. E. Neggers et al., “Target identification of small molecules using large-scale CRISPR-Cas mutagenesis scanning of essential genes,” NATURE COMMUNICATIONS, vol. 9, 2018.
@article{01H1Y107JN3JTNKNZRZW6TZXCB, abstract = {{Unraveling the mechanism of action and molecular target of small molecules remains a major challenge in drug discovery. While many cancer drugs target genetic vulnerabilities, loss-of-function screens fail to identify essential genes in drug mechanism of action. Here, we report CRISPRres, a CRISPR-Cas-based genetic screening approach to rapidly derive and identify drug resistance mutations in essential genes. It exploits the local genetic variation created by CRISPR-Cas-induced non-homologous end-joining (NHEJ) repair to generate a wide variety of functional in-frame mutations. Using large sgRNA tiling libraries and known drug-target pairs, we validate it as a target identification approach. We apply CRISPRres to the anticancer agent KPT-9274 and identify nicotinamide phosphoribosyltransferase (NAMPT) as its main target. These results present a powerful and simple genetic approach to create many protein variants that, in combination with positive selection, can be applied to reveal the cellular target of small-molecule inhibitors.}}, articleno = {{502}}, author = {{Neggers, Jasper Edgar and Kwanten, Bert and Dierckx, Tim and Noguchi, Hiroki and Voet, Arnout and Bral, Lotte and Minner, Kristien and Massant, Bob and Kint, Nicolas and Delforge, Michel and Vercruysse, Thomas and Baloglu, Erkan and Senapedis, William and Jacquemyn, Maarten and Daelemans, Dirk}}, issn = {{2041-1723}}, journal = {{NATURE COMMUNICATIONS}}, language = {{eng}}, pages = {{14}}, title = {{Target identification of small molecules using large-scale CRISPR-Cas mutagenesis scanning of essential genes}}, url = {{http://doi.org/10.1038/s41467-017-02349-8}}, volume = {{9}}, year = {{2018}}, }
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