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Next-generation sequencing to generate interactome datasets

2011; Nature Portfolio; Volume: 8; Issue: 6 Linguagem: Inglês

10.1038/nmeth.1597

1548-7105

Haiyuan Yu, Leah Tardivo, Stanley Tam, Evan Weiner, Fana Gebreab, Changyu Fan, Nenad Svrzikapa, Tomoko Hirozane-Kishikawa, Edward A. Rietman, Xinping Yang, Julie M. Sahalie, Kourosh Salehi‐Ashtiani, Tong Hao, Michael E. Cusick, David E. Hill, Frederick P. Roth, Pascal Braun, Marc Vidal,

Biotin and Related Studies

A combination of PCR stitching with next-generation sequencing results in Stitch-seq, a massively parallel method for interactome mapping. The approach is applied to human protein-protein interactions assayed in yeast two-hybrid screens. Next-generation sequencing has not been applied to protein-protein interactome network mapping so far because the association between the members of each interacting pair would not be maintained in en masse sequencing. We describe a massively parallel interactome-mapping pipeline, Stitch-seq, that combines PCR stitching with next-generation sequencing and used it to generate a new human interactome dataset. Stitch-seq is applicable to various interaction assays and should help expand interactome network mapping.