Prof. Viktor Stein ^{Centre for Synthetic Biology, Technical University Darmstadt}

Combinatorial Protein Engineering in the Context of the iFLinkC Framework
The iterative functional linker cloning (iFLinkC) framework provides a scalable, rule-based DNA assembly framework for constructing complex fusion proteins in a hierarchical and combinatorial fashion. iFLinkC has proven especially powerful in the context of combinatorial linker engineering which is turning out key in the construction of many high-performing protein switches and sensors. Successful examples include synthetic protease switches, bioluminescent and fluorescent protein sensors as well as recombinant nanopore assemblies. Finally, iFLinkC can be readily automated on a robotic platform.

Dr. Leon Kraus ^{Technical University Darmstadt}

Synthetic RNA Biology
RNA devices such as riboswitches, ribozymes, and biosensors have become essential tools in synthetic and molecular biology, leveraging RNA’s structural diversity and modularity to create programmable systems for sensing, gene regulation, and beyond. Aptamers, which bind targets with high affinity and specificity, are central to these technologies.

However, the selection of conformational-switching aptamers that are capable of driving these devices remains a challenge using traditional methods. We address this by developing integrated protocols for aptamer selection and engineering versatile devices from structure-switching RNAs. This has yielded multiple functional sequences, including a tobramycin aptamer that functions as a translation-regulating riboswitch, repressing gene expression by up to 18-fold.

A levofloxacin aptamer enabled a lateral flow assay platform that detects analytes via released capture oligonucleotides - a mechanism distinct from traditional direct-binding approaches. Finally, we engineered a new caffeine aptamer into functional hammerhead aptazymes in both yeast and mammalian cells via optofluidic screening. Integration with the iSpinach fluorogenic aptamer yielded a caffeine-responsive biosensor. These protocols expand the toolkit for RNA-based devices and open new avenues for diagnostics and synthetic biology applications.