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Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters

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Published on: November 26, 2013

A novel directed evolution platform for engineering chemically gated protein switches.

Luis A Vázquez-Rivera^1,2, Jiaqi Shen^1,3, Gwendolyn Shingles^1,3

¹Life Sciences Institute, University of Michigan, Ann Arbor, Michigan, USA. wenjwang@umich.edu.

Chemical Communications (Cambridge, England)

|March 3, 2026

Summary

We developed DuoSelect, a novel yeast surface selection platform, to efficiently identify chemical-dependent protein switches. This method combines positive and negative selections for improved signal-to-background ratios in a single step.

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Area of Science:

Biochemistry
Molecular Biology
Synthetic Biology

Background:

Chemically gated protein switches are crucial for controlling cellular functions.
Existing selection methods for protein switches can be inefficient.

Purpose of the Study:

To introduce DuoSelect, a new yeast surface selection platform.
To enable efficient selection of protein switches based on signal-to-background ratios.

Main Methods:

DuoSelect integrates positive and negative selection in one round of yeast sorting.
The platform was used to select two distinct chemical-dependent protein switches.

Main Results:

Successfully selected two novel chemical-dependent protein switches.
Demonstrated the utility of DuoSelect for high-throughput screening.
Showcased the application of selected protein switches in cellular assays.

Conclusions:

DuoSelect is an effective platform for selecting high-performance protein switches.
This platform accelerates the development of tools for chemical biology and synthetic biology.