FungalBraid 2.0: expanding the synthetic biology toolbox for the biotechnological exploitation of filamentous fungi

• DOI: 10.3389/fbioe.2023.1222812
• Publication Date: 2023-08-07T18:19:07Z
• Authors: Elena Moreno-Giménez; Mónica Gandía; Zara Sáez; Paloma Manzanares; Lynne Yenush; Diego Orzáez; Jose F. Marcos; Sandra Garrigues
• Keywords: Synthetic Biology; Cloning (programming); Selectable marker

Fungal synthetic biology is a rapidly expanding field that aims to optimize the biotechnological exploitation of fungi through generation standard, ready-to-use genetic elements, and universal syntax rules for contributory use by fungal research community. Recently, an increasing number toolkits have been developed applied filamentous fungi, which highlights relevance these organisms in biotechnology field. The FungalBraid (FB) modular cloning platform enables interchangeability DNA parts with GoldenBraid (GB) platform, designed plants, other systems are compatible standard Golden Gate syntax, uses binary pCAMBIA-derived vectors allow Agrobacterium tumefaciens-mediated transformation wide range species. In this study, we expanded original FB catalog adding 27 new were functionally validated vivo. Among resistance selection markers antibiotics phleomycin terbinafine, as well uridine-auxotrophic marker pyr4. We also used normalized luciferase reporter system validate several promoters, such PpkiA, P7760, Pef1α, PafpB constitutive PglaA, PamyB, PxlnA inducible promoters. Additionally, recently dCas9-regulated GB_SynP promoter collection orthogonal CRISPR activation (CRISPRa) plants has adapted system. general, expansion great interest scientific community since it increases possible interchangeable assemblies, exponentially possibilities studying, developing, exploiting fungi.