Journal
CELL
Volume 175, Issue 4, Pages 1141-+Publisher
CELL PRESS
DOI: 10.1016/j.cell.2018.09.022
Keywords
-
Categories
Funding
- NIH [R01AI113221, R33CA182377, U19AI128949, S10OD023547]
- Belgian American Educational Foundation
- OCRFA Liz Tilberis Award
- NATIONAL CANCER INSTITUTE [R01CA154947, R33CA182377] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [R01AI113221, U24AI118644, U19AI128949] Funding Source: NIH RePORTER
- OFFICE OF THE DIRECTOR, NATIONAL INSTITUTES OF HEALTH [S10OD023547] Funding Source: NIH RePORTER
Ask authors/readers for more resources
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 analyzedmultiple 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 proteinlevel phenotyping of 100s of genes with single-cell resolution.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available