Pooled‐matrix protein interaction screens using Barcode Fusion Genetics
View/ Open
Author
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
Tyagi, Tanya
Sheykhkarimli, Dayag
Roth, Jonathan F
Wong, Cassandra
Musa, Louai
Snider, Jamie
Liu, Yi‐Chun
Yu, Haiyuan
Braun, Pascal
Stagljar, Igor
Hao, Tong
Pelletier, Laurence
Aloy, Patrick
Roth, Frederick P
Note: Order does not necessarily reflect citation order of authors.
Published Version
https://doi.org/10.15252/msb.20156660Metadata
Show full item recordCitation
Yachie, N., E. Petsalaki, J. C. Mellor, J. Weile, Y. Jacob, M. Verby, S. B. Ozturk, et al. 2016. “Pooled‐matrix protein interaction screens using Barcode Fusion Genetics.” Molecular Systems Biology 12 (4): 863. doi:10.15252/msb.20156660. http://dx.doi.org/10.15252/msb.20156660.Abstract
Abstract 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.Other Sources
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4848762/pdf/Terms of Use
This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAACitable link to this page
http://nrs.harvard.edu/urn-3:HUL.InstRepos:27320197
Collections
- HMS Scholarly Articles [17922]
Contact administrator regarding this item (to report mistakes or request changes)