Pooled-matrix protein interaction screens using Barcode Fusion Genetics

High-throughput binary protein interaction mapping is continuing to extend our understanding of cellular function and disease mechanisms. However, we remain one or two orders of magnitude away from a complete interaction map for humans and other major model organisms. Completion will require screening at substantially larger scales with many complementary assays, requiring further efficiency gains in proteome-scale interaction mapping. Here, we report Barcode Fusion Genetics-Yeast Two-Hybrid (BFG-Y2H), by which a full matrix of protein pairs can be screened in a single multiplexed strain pool. BFG-Y2H uses Cre recombination to fuse DNA barcodes from distinct plasmids, generating chimeric protein-pair barcodes that can be quantified via next-generation sequencing. We applied BFG-Y2H to four different matrices ranging in scale from ~25 K to 2.5 M protein pairs. The results show that BFG-Y2H increases the efficiency of protein matrix screening, with quality that is on par with state-of-the-art Y2H methods.

Source: Molecular Systems Biology - April 21, 2016 Category: Molecular Biology Authors: Yachie, N., Petsalaki, E., Mellor, J. C., Weile, J., Jacob, Y., Verby, M., Ozturk, S. B., Li, S., Cote, A. G., Mosca, R., Knapp, J. J., Ko, M., Yu, A., Gebbia, M., Sahni, N., Yi, S., Tyagi, T., Sheykhkarimli, D., Roth, J. F., Wong, C., Musa, L., Snider, J Tags: Methods & Resources, Network Biology Articles Source Type: research