PASTMUS: mapping functional elements at single amino acid resolution in human cells

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.

确定感兴趣蛋白质的功能元件对于实现对其机制的理解非常重要。然而，评估给定蛋白质的每一个氨基酸与其功能重要性的相关性仍然很麻烦。在此，我们报告了一种策略，即从CRISPR平铺诱变筛选（PASTMUS）中解析断裂的DNA序列，它提供了一种简化的工作流程和一个生物信息学管道，以便在其天然生物学背景下识别蛋白质的关键氨基酸。利用这种方法，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，并获得了它们相应的氨基酸分辨率的功能图谱。