CRISPR-based peptide library display and programmable microarray self-assembly for rapid quantitative protein binding assays.
Mol Cell
; 81(17): 3650-3658.e5, 2021 09 02.
Article
in English
| MEDLINE | ID: covidwho-1356368
ABSTRACT
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.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
RNA, Guide, Kinetoplastida
/
Peptide Library
/
Gene Editing
/
Epitopes
Type of study:
Diagnostic study
/
Randomized controlled trials
Limits:
Humans
Language:
English
Journal:
Mol Cell
Journal subject:
Molecular Biology
Year:
2021
Document Type:
Article
Affiliation country:
J.molcel.2021.07.027
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