CRISPR-inspired systems have been extensively developed for applications in genome editing and nucleic acid detection. Here, we introduce a CRISPR-based peptide display technology to facilitate customized, high-throughput in vitro protein interaction studies. We show that bespoke peptide libraries fused to catalytically inactive Cas9 (dCas9) and barcoded with unique single guide RNA (sgRNA) molecules self-assemble from a single mixed pool to programmable positions on a DNA microarray surface for rapid, multiplexed binding assays. We develop dCas9-displayed saturation mutagenesis libraries to characterize antibody-epitope binding for a commercial anti-FLAG monoclonal antibody and human serum antibodies. We also show that our platform can be used for viral epitope mapping and exhibits promise as a multiplexed diagnostics tool. Our CRISPR-based peptide display platform and the principles of complex library self-assembly using dCas9 could be adapted for rapid interrogation of varied customized protein libraries or biological materials assembly using DNA scaffolding.

受 CRISPR 启发的系统已被广泛开发用于基因组编辑和核酸检测中的应用。在这里，我们介绍了一种基于 CRISPR 的肽展示技术，以促进定制的、高通量的体外蛋白质相互作用研究。我们展示了与催化失活的 Cas9 (dCas9) 融合并用独特的单向导 RNA (sgRNA) 分子标记的定制肽库，从单个混合池自组装到 DNA 微阵列表面上的可编程位置，用于快速、多重结合测定。我们开发了 dCas9 展示的饱和诱变文库，以表征商业抗 FLAG 单克隆抗体和人血清抗体的抗体表位结合。我们还表明，我们的平台可用于病毒表位作图，并有望成为一种多重诊断工具。我们基于 CRISPR 的肽展示平台和使用 dCas9 的复杂文库自组装原理可适用于使用 DNA 支架快速查询各种定制蛋白质文库或生物材料组装。