Project Overview

Many useful proteins are difficult to express or use because they misfold, aggregate, or lose activity. This creates problems for protein production, storage, and application in biotechnology.

Our project explores whether intrinsically disordered protein, or IDP, tags can help target proteins remain soluble and functional. We are combining computational design with experimental selection to evolve better tags.

Our Rationale

IDPs are flexible proteins that do not adopt one fixed structure. This flexibility may help them act as solubility tags, protective partners, or stress-tolerance elements.

Instead of redesigning each difficult protein individually, we want to test whether short transferable tags can improve protein behaviour across different systems.

Project Goals

• Design libraries of candidate IDP tags.
• Test whether these tags improve target protein performance.
• Identify sequence features linked to improved solubility or stability.
• Use selection results to guide the next round of design.
• Document our workflow so future teams can build on it.

Technical Workflow

Our workflow begins with known and de novo IDP-inspired sequences. These are analysed computationally, filtered, converted into DNA libraries, cloned into expression systems, and tested experimentally.

A key part of the project is library design. We need enough diversity to find improved tags, but not so much degeneracy that most generated sequences lose useful sequence relationships.

Experimental Plan

We are developing two main experimental branches. The first uses beta-lactamase as a selectable target protein, where improved tag performance may allow cells to survive higher antibiotic stress.

The second explores fluorescent or enzymatic readouts, including tardigrade-inspired protection systems and LDH-based activity assays.

Why Directed Evolution?

Protein solubility is difficult to predict perfectly from sequence alone. Directed evolution lets us generate variants, test them experimentally, and use the results to improve the next generation of designs.

Current Direction

We are currently refining our library design and experimental workflow. In particular, we are comparing broad degenerate libraries, reduced-degeneracy libraries, and error-prone PCR as possible routes for generating useful tag variants.

Latest Update

What to Read Next

Visit the archive for meeting updates, design notes, experimental plans, and longer project write-ups as the wiki grows.