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.
['Yachie|Nozomu|N|', 'Petsalaki|Evangelia|E|', 'Mellor|Joseph C|JC|', 'Weile|Jochen|J|', 'Jacob|Yves|Y|', 'Verby|Marta|M|', 'Ozturk|Sedide B|SB|', 'Li|Siyang|S|', 'Cote|Atina G|AG|', 'Mosca|Roberto|R|', 'Knapp|Jennifer J|JJ|', 'Ko|Minjeong|M|', 'Yu|Analyn|A|', 'Gebbia|Marinella|M|', 'Sahni|Nidhi|N|', 'Yi|Song|S|', 'Tyagi|Tanya|T|', 'Sheykhkarimli|Dayag|D|', 'Roth|Jonathan F|JF|', 'Wong|Cassandra|C|', 'Musa|Louai|L|', 'Snider|Jamie|J|', 'Liu|Yi-Chun|YC|', 'Yu|Haiyuan|H|', 'Braun|Pascal|P|', 'Stagljar|Igor|I|', 'Hao|Tong|T|', 'Calderwood|Michael A|MA|http://orcid.org/0000-0001-6475-1418', 'Pelletier|Laurence|L|', 'Aloy|Patrick|P|', 'Hill|David E|DE|http://orcid.org/0000-0001-5192-0921', 'Vidal|Marc|M|http://orcid.org/0000-0003-3391-5410', 'Roth|Frederick P|FP|']
D018385:Centrosome D002877:Chromosomes, Human D015723:Gene Library D059014:High-Throughput Nucleotide Sequencing D006801:Humans D011485:Protein Binding D025941:Protein Interaction Mapping D020543:Proteome D012441:Saccharomyces cerevisiae D020798:Two-Hybrid System Techniques
protein interaction DNA barcode interactome next‐generation sequencing yeast two‐hybrid