Identification of functional elements for a protein of interest is important for achieving a mechanistic understanding. However, it remains cumbersome to assess each and every amino acid of a given protein in relevance to its functional significance. Here, we report a strategy, PArsing fragmented DNA Sequences from CRISPR Tiling MUtagenesis Screening (PASTMUS), which provides a streamlined workflow and a bioinformatics pipeline to identify critical amino acids of proteins in their native biological contexts. Using this approach, we map six proteins—three bacterial toxin receptors and three cancer drug targets, and acquire their corresponding functional maps at amino acid resolution.

中文翻译：

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PASTMUS：在人类细胞中以单一氨基酸分辨率绘制功能元件

鉴定感兴趣蛋白质的功能元件对于实现机理理解很重要。然而，评估给定蛋白质的每个氨基酸与其功能意义的相关性仍然很麻烦。在这里，我们报告了一种策略，即从 CRISPR 平铺突变筛选 (PASTMUS) 中解析片段化 DNA 序列，该策略提供了简化的工作流程和生物信息学管道，以识别蛋白质在其天然生物环境中的关键氨基酸。使用这种方法，我们绘制了六种蛋白质——三种细菌毒素受体和三种癌症药物靶点，并以氨基酸分辨率获得它们相应的功能图谱。