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CRISPR-based peptide library display and programmable microarray self-assembly for rapid quantitative protein binding assays

Mol Cell. 2021-08; 
Karl W Barber, Ellen Shrock, Stephen J Elledge
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Custom DNA/RNA Oligos … using dCas9 could be adapted for rapid interrogation of varied customized protein libraries or biological materials assembly using DNA … Oligonucleotide microarrays were ordered from Customarray (GenScript) containing four subarrays each with 2,240 individual features. Each … Get A Quote

摘要

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... More

关键词

CRISPR, Cas9, FLAG, SARS-CoV-2, antibody binding, influenza A, protein display, protein microarray, protein-protein interactions, self-assembly