CasPlay provides a gRNA-barcoded CRISPR-based display platform for antibody repertoire profiling

Karl W Barber; Ellen Shrock; Stephen J Elledge

doi:10.1016/j.crmeth.2022.100318

CasPlay provides a gRNA-barcoded CRISPR-based display platform for antibody repertoire profiling

Cell Rep Methods. 2022 Oct 17;2(10):100318. doi: 10.1016/j.crmeth.2022.100318. eCollection 2022 Oct 24.

Authors

Karl W Barber^{1

2}, Ellen Shrock^{1

2}, Stephen J Elledge^{1

2}

Affiliations

¹ Division of Genetics, Brigham and Women's Hospital, Howard Hughes Medical Institute, Boston, MA 02115, USA.
² Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.

Abstract

Protein display technologies link proteins to distinct nucleic acid sequences (barcodes), enabling multiplexed protein assays via DNA sequencing. Here, we develop Cas9 display (CasPlay) to interrogate customized peptide libraries fused to catalytically inactive Cas9 (dCas9) by sequencing the guide RNA (gRNA) barcodes associated with each peptide. We first confirm the ability of CasPlay to characterize antibody epitopes by recovering a known binding motif for a monoclonal anti-FLAG antibody. We then use a CasPlay library tiling the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) proteome to evaluate vaccine-induced antibody reactivities. Using a peptide library representing the human virome, we demonstrate the ability of CasPlay to identify epitopes across many viruses from microliters of patient serum. Our results suggest that CasPlay is a viable strategy for customized protein interaction studies from highly complex libraries and could provide an alternative to phage display technologies.

Keywords: CRISPR; Cas9; antibody binding; biotechnology; peptide display; protein display.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Antibodies
COVID-19*
Epitopes / chemistry
Humans
Peptide Library
SARS-CoV-2 / genetics

Substances

Peptide Library
Antibodies
Epitopes