Week 3 HW: Cell-Free Reaction Composition

Week 3 Homework – Designing Cell-Free Reaction Compositions

Overview

I designed cell-free protein synthesis (CFPS) reaction compositions for three proteins: sfGFP, PETase, and Reteplase using the coopetition.htgaa.org platform.

Compositions Submitted

1. sfGFP

• Type: Reducing condition
• Energy source: PEP-based
• Comments: Simple PEP-based composition based on reference plate.

2. PETase

• Type: Reducing condition
• Energy source: PEP-based
• Comments: Similar to sfGFP but suitable for an enzyme with one disulfide bond.

3. Reteplase

• Type: Oxidizing (DSB) condition
• Special reagents: GSSG, GSH, and DsbC
• Comments: Oxidizing composition to support formation of 9 disulfide bonds.

Community Reagent

I suggested Trehalose as a community reagent.
Reason: It acts as a protein stabilizer and can improve functional yield, especially for difficult proteins like Reteplase.

AI Note

I used Grok to help understand the requirements and structure the designs. Final decisions are my own.