Long-read sequencing to detect full-length protein-protein interactions
Listed in
This article is not in any list yet, why not save it to one of your lists.Abstract
Given the increased predictions on interactome size and demand for protein function information, methods for detecting protein-protein interactions remain a significant development area. The all-vs.-all sequencing (AVA-Seq) method utilizes a convergent fusion plasmid design to make two-hybrid technology amenable to next-generation sequencing. Here, we further innovate to take advantage of synthetic DNA technologies and Oxford Nanopore Technologies long-read sequencing improvements to allow us to determine full-length protein-protein interactions. Here, using this approach we recovered 159 protein-protein interactions from a set of 57 human proteins using multiple forms of validation. Further, when referencing a human gold standard set of interactions, eight full-length protein-protein interactions were recovered from an expected 28 interaction pairs (28.6%), a typical recovery rate for two-hybrid technologies. The AVA-Seq, in combination with the ease of synthetic DNA production and the MinION platform, offers a low-cost, high-throughput alternative for determining protein-protein interactions, which can be utilized in research labs at all stages.
Key Points
First application of long-read sequencing for full-length protein-protein interaction studies.
The recovery rate of the AVA-Seq method using full-length proteins is on par with other leading methods.
Advances in synthetic biology and sequencing technologies make full-length protein interactomes affordable and accessible.
