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Front Genet. 2015 Dec 17;6:346. doi: 10.3389/fgene.2015.00346. eCollection 2015.

Next-Generation Sequencing for Binary Protein-Protein Interactions.

Author information

1
Next Interactions, Inc., Richmond CA, USA.
2
BioInfoRx, Inc., Madison WI, USA.
3
Next Interactions, Inc., RichmondCA, USA; Regenerative Sciences Institute, SunnyvaleCA, USA.
4
Department of Plant Biology, University of California, Davis, Davis CA, USA.
5
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley CA, USA.

Abstract

The yeast two-hybrid (Y2H) system exploits host cell genetics in order to display binary protein-protein interactions (PPIs) via defined and selectable phenotypes. Numerous improvements have been made to this method, adapting the screening principle for diverse applications, including drug discovery and the scale-up for proteome wide interaction screens in human and other organisms. Here we discuss a systematic workflow and analysis scheme for screening data generated by Y2H and related assays that includes high-throughput selection procedures, readout of comprehensive results via next-generation sequencing (NGS), and the interpretation of interaction data via quantitative statistics. The novel assays and tools will serve the broader scientific community to harness the power of NGS technology to address PPI networks in health and disease. We discuss examples of how this next-generation platform can be applied to address specific questions in diverse fields of biology and medicine.

KEYWORDS:

interactome mapping; next-generation sequencing; protein–protein interactions; quantitative interaction profiles; yeast two-hybrid

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