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Protein Barcodes Enable High-Dimensional Single-Cell CRISPR Screens.
- Source :
-
Cell [Cell] 2018 Nov 01; Vol. 175 (4), pp. 1141-1155.e16. Date of Electronic Publication: 2018 Oct 18. - Publication Year :
- 2018
-
Abstract
- CRISPR pools are being widely employed to identify gene functions. However, current technology, which utilizes DNA as barcodes, permits limited phenotyping and bulk-cell resolution. To enable novel screening capabilities, we developed a barcoding system operating at the protein level. We synthesized modules encoding triplet combinations of linear epitopes to generate >100 unique protein barcodes (Pro-Codes). Pro-Code-expressing vectors were introduced into cells and analyzed by CyTOF mass cytometry. Using just 14 antibodies, we detected 364 Pro-Code populations; establishing the largest set of protein-based reporters. By pairing each Pro-Code with a different CRISPR, we simultaneously analyzed multiple phenotypic markers, including phospho-signaling, on dozens of knockouts. Pro-Code/CRISPR screens found two interferon-stimulated genes, the immunoproteasome component Psmb8 and a chaperone Rtp4, are important for antigen-dependent immune editing of cancer cells and identified Socs1 as a negative regulator of Pd-l1. The Pro-Code technology enables simultaneous high-dimensional protein-level phenotyping of 100s of genes with single-cell resolution.<br /> (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Subjects :
- Animals
Epitopes chemistry
Epitopes classification
Epitopes genetics
HEK293 Cells
Humans
Immunophenotyping methods
Jurkat Cells
Mice, Inbred BALB C
Proteome chemistry
Proteome classification
Proteome genetics
THP-1 Cells
CRISPR-Cas Systems
Flow Cytometry methods
Genomics methods
Mass Spectrometry methods
Single-Cell Analysis methods
Subjects
Details
- Language :
- English
- ISSN :
- 1097-4172
- Volume :
- 175
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Cell
- Publication Type :
- Academic Journal
- Accession number :
- 30343902
- Full Text :
- https://doi.org/10.1016/j.cell.2018.09.022