Invitris’ Post

🎉 Onto the last post in this lightning series! This time, let's take a closer look at an example of toxic protein expression using a cell-free system. 𝘋𝘪𝘴𝘤𝘭𝘢𝘪𝘮𝘦𝘳: 𝘛𝘩𝘪𝘴 𝘴𝘵𝘶𝘥𝘺 𝘪𝘴 𝘢𝘯 𝘪𝘯𝘥𝘦𝘱𝘦𝘯𝘥𝘦𝘯𝘵 𝘸𝘰𝘳𝘬 𝘣𝘺 𝘙𝘢𝘮𝘮 𝘦𝘵 𝘢𝘭. 𝘢𝘯𝘥 𝘪𝘴 𝘯𝘰𝘵 𝘳𝘦𝘭𝘢𝘵𝘦𝘥 𝘵𝘰 𝘐𝘯𝘷𝘪𝘵𝘳𝘪𝘴. 𝗥𝗲𝘃𝗼𝗹𝘂𝘁𝗶𝗼𝗻𝗶𝘇𝗶𝗻𝗴 𝘁𝗼𝘅𝗶𝗻 𝗿𝗲𝘀𝗲𝗮𝗿𝗰𝗵: 𝗦𝗵𝗶𝗴𝗮 𝘁𝗼𝘅𝗶𝗻 𝗲𝘅𝗽𝗿𝗲𝘀𝘀𝗶𝗼𝗻 𝘂𝘀𝗶𝗻𝗴 𝗰𝗲𝗹𝗹-𝗳𝗿𝗲𝗲 𝘀𝘆𝘀𝘁𝗲𝗺𝘀 🧬 Shiga toxins (Stx) belong to a group of AB5 toxins, characterized by a catalytic A-subunit that targets eukaryotic ribosomes and a pentameric B-subunit. With nearly 3 million cases of Stx-related illnesses reported annually, studying their mechanism of action and developing new treatments is crucial. 🔬 A recent study explored the potential of Cell-Free Protein Synthesis (CFPS) for expressing Shiga toxin, using E. coli and CHO-derived cell extracts. This research, by Franziska Ramm and colleagues, published in Scientific Reports, highlights both the promise and challenges of CFPS in toxin biology, offering a novel approach to studying dangerous proteins in a safer environment. 📊 Key Findings & Impact: ✅ Safe expression: CFPS enabled the synthesis of Shiga toxin without the need for live cells. ✅ Functional validation: The study confirmed the biological activity of CFPS-produced Shiga toxins through ribosome-inhibition assays and OPP assays in HeLa cells. ⚠️ Challenges in yield: While successful, CFPS-generated toxin levels in the CHO-based system were lower than in 𝘌.𝘤𝘰𝘭𝘪 CFPS and traditional cell-based methods, highlighting the need for further optimization. This study underscores the growing role of CFPS in synthetic biology, particularly for expressing challenging proteins that are difficult to work with in live-cell systems. 🔗 Interested to learn more? Read the full paper: Ramm, F., Kaser, D., König, I. et al. Synthesis of biologically active Shiga toxins in cell-free systems. Sci Rep 14, 6043 (2024). https://lnkd.in/dajsS99q 💡 What are your thoughts on the future of CFPS in toxin research? Let’s discuss! 👇 #Invitris #SynBio #CellFree