Services

Next Generation Sequencing
Yeast Two-Hybrid Screening Services

We offer a suite of services comprising protein-protein interaction (PPI) screens, protein engineering and functional assays. Our clients are in pharma, biotech and academic research.

We use Next-Generation Sequencing (NGS) as the readout, resulting in more complete datasets. Our thoroughly redesigned Y2H system components provide more quantitative and precise measurements of binding partner interactions than conventional Y2H assays.  Our platform technology enables us to decipher the full coverage of protein-protein interactions (PPIs). These discoveries have been applied to problems in human health, energy, food security and sustainability.

Conventional Yeast Two-Hybrid Screening

Conventional Yeast Two-Hybrid Screening

Conventional Yeast Two-Hybrid Service offered for clients who need simple solutions for their project, and are on a restricted budget. The traditional Y2H uses the Sanger (1st generation or capillary) sequencing as a readout. It is simple and cheap but has some drawbacks compared to the NGS-Y2H, which uses the next generation sequencing as readout.

For example, you want to screen a known protein as a bait against a library of prey proteins generated from the tissue of your interest.

Conventional Yeast Two-Hybrid Screening
Side View, Yeast Inoculation Done By Scientist

One-on-one Protein Interaction Test (OPIT)

The one-on-one protein interaction test is a simple bait to prey interaction test to check if two given proteins are interacting (no screening of a library). This can be done with a visual readout (qualitative and quantitative growth assays). (References: Shuguang et al , PNAS, 2018, Nicolina Sostaric et al,  Molecular & Cellular Proteomics, 2018)

For example, you already know that two proteins are present in a pathway, but there is no evidence so far for interaction. You want to confirm it. Or you want to study mutation or variants of the protein in 1:1 assays for visualization.

Basic NGS-Yeast Two-Hybrid Screening

Basic NGS-Yeast Two-Hybrid Screening

NGS coupled Yeast Two-Hybrid Screening service for finding new protein-protein interactions, for those who want an advanced and comprehensive readout for their screens.  With NGS, the screening is more reliable, more precise than the conventional Y2H screening using first-generation capillary sequencing. Expect up to 10x more hits.

For example, you are interested in a protein or a protein domain and you need to get an idea of what its function is. We will clone the gene of interest into a bait vector and screen this gene with a cDNA library from a relevant source (cells, tissue). You will get then a list of genes that encode potential interactors for your bait proteins. Pathway enrichment analysis highlights the biological roles of the bait protein.

Basic NGS-Yeast Two-Hybrid Screening
Premium NGS Yeast-based Two-Hybrid Screening

Premium NGS-Yeast Based Two-Hybrid Screening

Comprehensive interaction profiling and comparison to controls using quantitative statistics. NGS coupled Y2H service uses background controls and proprietary bioinformatics analysis to exclude false positives a priori from the results. We apply three assay repeats (triplicates) to enable statistics. With this procedure, we are able to cross-compare interaction patterns between different bait proteins.

For example, you have a mutation in an oncogene that is supposed to underlie a role in cancer. We will generate bait constructs for mutant and wild-type proteins and then use them to screen a cDNA library of your choice side by side. All screens are done in triplicates to generate statistical power to determine bait specific enrichments.

PPI Mapping with NGS-Yeast Two-Hybrid

PPI Mapping with NGS-Yeast Two-Hybrid

NGS coupled Y2H precision mapping service and high-level bioinformatics to characterize protein-protein interaction sites at amino acid resolution.

For example, you know that an oncogene binds to its effector in the cancer pathway. This interaction was already identified and published by other researchers. However, there is no good data or conflicting data on the exact position of the binding sequence, i.e. which residues you need to focus on for a mutagenesis experiment. We will generate a high-complexity library of fragments saturating the target sequence. We will determine the exact binding site using Y2H-NGS technology.

Final Image for PM lrg cr
HTP Screening with  NGS-Yeast Two-Hybrid

HTP Screening with  NGS-Yeast Two-Hybrid

The NGS based Y2H can be done in a high throughput manner (>100 baits) with many customized experimental and computational procedures, thus greatly diminishing the cost per screen.

For example, you want to get an overview of a disease pathway or the disease mechanism of a pathogen that launches multiple effector proteins to thoroughly modulate the physiology and immune response of its host. We will perform a parallelized screening procedure with all effector proteins as baits vs a host cell cDNA library. We can then score hits and pathway enrichments to generate an interaction network and to elucidate the disease mechanism.

Services

Compound / Drug Screening with NGS Yeast Two-Hybrid

Taking advantage of the improved screening format that is enabled by NGS-Y2H procedure, we are able to directly assess drug effects on PPIs, either looking for disruption of already known target interactions, or for drug-induced complex assemblies. Tests can be done in different formats from 1:1 assays to comparative and interaction profiling.

For example, you would like to find the best out of 1-10 compounds that would disable the interaction of several known proteins with a target in a cancer pathway. We can screen interactions in the presence or absence of a large number of compounds in a small scale or in a high throughput manner. Drug dependent interaction profiles are then scored.

Compound / Drug Screening with NGS Yeast Two-Hybrid

Receive more information about next generation yeast two-hybrid screening services.

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Contact us to schedule a consultation and we will answer all your questions.