| First Author | Wroblewska A | Year | 2018 |
| Journal | Cell | Volume | 175 |
| Issue | 4 | Pages | 1141-1155.e16 |
| PubMed ID | 30343902 | Mgi Jnum | J:355263 |
| Mgi Id | MGI:6259377 | Doi | 10.1016/j.cell.2018.09.022 |
| Citation | Wroblewska A, et al. (2018) Protein Barcodes Enable High-Dimensional Single-Cell CRISPR Screens. Cell 175(4):1141-1155.e16 |
| abstractText | 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. |
