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

被引：18

作者：

Barber, Karl W. ^{[1
,2
]}

Shrock, Ellen ^{[1
,2
]}

Elledge, Stephen J. ^{[1
,2
]}

机构：

[1] Brigham & Womens Hosp, Howard Hughes Med Inst, Div Genet, Boston, MA 02115 USA

[2] Harvard Med Sch, Dept Genet, Boston, MA 02115 USA

来源：

MOLECULAR CELL | 2021年 / 81卷 / 17期

基金：

美国国家科学基金会;

关键词：

SEQUENCE LOGOS; PHAGE DISPLAY; SPECIFICITY; REPRESSION; SCALE;

D O I：

10.1016/j.molcel.2021.07.027

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

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 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.

引用

页码：3650 / +

页数：15

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