Open AccessPooled‐matrix protein interaction screens using Barcode Fusion Genetics
Author(s) -
Yachie Nozomu,
Petsalaki Evangelia,
Mellor Joseph C,
Weile Jochen,
Jacob Yves,
Verby Marta,
Ozturk Sedide B,
Li Siyang,
Cote Atina G,
Mosca Roberto,
Knapp Jennifer J,
Ko Minjeong,
Yu Analyn,
Gebbia Marinella,
Sahni Nidhi,
Yi Song,
Tyagi Tanya,
Sheykhkarimli Dayag,
Roth Jonathan F,
Wong Cassandra,
Musa Louai,
Snider Jamie,
Liu YiChun,
Yu Haiyuan,
Braun Pascal,
Stagljar Igor,
Hao Tong,
Calderwood Michael A,
Pelletier Laurence,
Aloy Patrick,
Hill David E,
Vidal Marc,
Roth Frederick P
Publication year - 2016
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.20156660
Subject(s) - barcode , biology , computational biology , genetics , fusion protein , computer science , gene , recombinant dna , operating system
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.