清脆的
计算生物学
生物
突变
药物发现
基因
基因组编辑
遗传学
Cas9
外显子
基因组
突变
生物信息学
作者
Junwei Shi,Eric Wang,Joseph P. Milazzo,Zihua Wang,Justin B. Kinney,Christopher R. Vakoc
摘要
Cancer drug targets are identified by CRISPR-based screens that knock out functional protein domains. CRISPR-Cas9 genome editing technology holds great promise for discovering therapeutic targets in cancer and other diseases. Current screening strategies target CRISPR-Cas9–induced mutations to the 5′ exons of candidate genes1,2,3,4,5, but this approach often produces in-frame variants that retain functionality, which can obscure even strong genetic dependencies. Here we overcome this limitation by targeting CRISPR-Cas9 mutagenesis to exons encoding functional protein domains. This generates a higher proportion of null mutations and substantially increases the potency of negative selection. We also show that the magnitude of negative selection can be used to infer the functional importance of individual protein domains of interest. A screen of 192 chromatin regulatory domains in murine acute myeloid leukemia cells identifies six known drug targets and 19 additional dependencies. A broader application of this approach may allow comprehensive identification of protein domains that sustain cancer cells and are suitable for drug targeting.
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