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

Emily K BowmanJames M WagnerShuo-Fu YuanMatthew DeanerClaire M PalmerSimon D'OelsnitzLauren T CordovaXin LiFrank F CraigHal S Alper
Published in: Proceedings of the National Academy of Sciences of the United States of America (2022)
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.
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