(174bl) Development of a Synthetic Biology Toolkit for Filamentous Fungi to Program Engineered Living Materials

Biomaterials derived from filamentous fungi have received growing interest as these organisms form strong mycelial structures, are resilient to environmental stresses, and can grow on cheap substrates. The application of synthetic biology to these fungi could enable the creation of engineered living materials (ELMs) capable of tailored functions such as sensing and responding to environmental stimuli. Although filamentous fungi play an increasingly important role in biomanufacturing, genetic tools suitable for these strains (especially among basidiomycetes) are significantly underdeveloped compared to those for bacteria and yeasts. We therefore aim to significantly expand the synthetic biology tools available for ELM-relevant fungal strains. An agrobacterium-mediated transformat­­ion protocol is first optimized for multiple ELM-relevant fungi species by screening various agrobacterium strains, selection markers, and transformation conditions. Libraries of genetic parts (e.g., inducible promoters, constitutive promoters, terminators) are then constructed and tested. Development of a robust synthetic biology toolkit applicable in a broad range of filamentous fungi is a critical step in engineering living materials capable of performing complex functions.