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- Title
De novo identification of essential protein domains from CRISPR-Cas9 tiling-sgRNA knockout screens.
- Authors
He, Wei; Zhang, Liang; Villarreal, Oscar D.; Fu, Rongjie; Bedford, Ella; Dou, Jingzhuang; Patel, Anish Y.; Bedford, Mark T.; Shi, Xiaobing; Chen, Taiping; Bartholomew, Blaine; Xu, Han
- Abstract
High-throughput CRISPR-Cas9 knockout screens using a tiling-sgRNA design permit in situ evaluation of protein domain function. Here, to facilitate de novo identification of essential protein domains from such screens, we propose ProTiler, a computational method for the robust mapping of CRISPR knockout hyper-sensitive (CKHS) regions, which refer to the protein regions associated with a strong sgRNA dropout effect in the screens. Applied to a published CRISPR tiling screen dataset, ProTiler identifies 175 CKHS regions in 83 proteins. Of these CKHS regions, more than 80% overlap with annotated Pfam domains, including all of the 15 known drug targets in the dataset. ProTiler also reveals unannotated essential domains, including the N-terminus of the SWI/SNF subunit SMARCB1, which is validated experimentally. Surprisingly, the CKHS regions are negatively correlated with phosphorylation and acetylation sites, suggesting that protein domains and post-translational modification sites have distinct sensitivities to CRISPR-Cas9 mediated amino acids loss. Tiling-sgRNA designs allow the in situ evaluation of protein domain functions. Here the authors present ProTiler - a computational method to predict CRISPR knockout hyper-sensitive regions, revealing previously unannotated domains.
- Subjects
PROTEIN domains; CRISPRS; NON-coding RNA; TARGETED drug delivery; PHOSPHORYLATION
- Publication
Nature Communications, 2019, Vol 10, Issue 1, pN.PAG
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-019-12489-8