"EPICode, a multiplexed epitope barcoding strategy that enables dynamic cellular phenotypic screens"

BME 500 Seminar: Markus Covert, Ph.D. Bioengineering, Stanford University

WHERE: Virtual

WHEN: December 9, 2021 4:00 pm-5:00 pmADD TO CALENDAR

Pooled genetic libraries have improved screening throughput for mapping genotypes to phenotypes. However, selectable phenotypes are limiting, restricting screening to outcomes with a low spatiotemporal resolution. Here, we integrated live-cell imaging with pooled library-based screening. To enable intracellular multiplexing, we developed a method, called EPICode (Epitope-Phenotype Immunofluorescence barCode), that uses a combination of short epitopes to facilitate optical screening. By using the subcellular localization of the barcode as an additional channel, our method exponentially increases multiplexing capacity. Thus, after using live-cell microscopy to characterize a phenotype of interest, subjected to sequential stimulatory/inhibitory manipulations, the genotype of each cell in the population can be identified. To demonstrate applicability, we developed a live-cell PKA kinase translocation reporter with improved sensitivity and specificity. The use of epitopes as fluorescent barcodes introduces a scalable strategy for high-throughput screening broadly applicable to protein engineering and drug discovery settings where image-based phenotyping is desired.
Details:
DATE: Thursday, December 9, 2021
TIME: 4:00-5:00 pm
ZOOM LINK: https://umich.zoom.us/j/97723483179