Sorting for secreted molecule production using a biosensor-in-microdroplet approach

Sorting large libraries of cells for improved small molecule secretion is throughput limited. Here, we combine producer/secretor cell libraries with whole-cell biosensors using a microfluidic-based screening workflow. This approach enables a mix-and-match capability using off-the-shelf biosensors through either coencapsulation or pico-injection. We demonstrate the cell type and library agnostic nature of this workflow by utilizing single-guide RNA, transposon, and ethyl-methyl sulfonate mutagenesis libraries across three distinct microbes ( Escherichia coli , Saccharomyces cerevisiae , and Yarrowia lipolytica ), biosensors from two organisms ( E. coli and S. cerevisiae ), and three products (triacetic acid lactone, naringenin, and L-DOPA) to identify targets improving production/secretion.