Microfluidic droplet-based high-throughput screening of filamentous fungi

Filamentous fungi are an extremely essential source of industrial enzymes due to their capacity to secrete large quantities of proteins. Creating knockout mutants in filamentous fungi is typically needed to understand the functions of their genes as well as to utilize such information to develop phenotypes with improved capabilities. However, the process of identifying a true knockout mutant requires screening of a large number of genetic transformants. Here, we developed a droplet microfluidics-based single-cell-resolution high-throughput screening strategy to select and isolate filamentous fungal transformants that are true knockouts mutants. The successful proof-of-concept assays demonstrated here make this platform ready for high-throughput filamentous fungal screening to facilitate filamentous fungi research and development of them as microbial bioproduction platforms.