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 th...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2021-09, Vol.118 (36), p.1-10, Article 2106818118 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Bowman, Emily K. Wagner, James M. Yuan, Shuo-Fu Deaner, Matthew Palmer, Claire M. D’Oelsnitz, Simon Cordova, Lauren Li, Xin Craig, Frank F. Alper, Hal S. |
| description | 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. |
| doi_str_mv | 10.1073/pnas.2106818118 |
| format | Article |
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| subjects | Biological Sciences Biosensing Techniques Escherichia coli - genetics Escherichia coli - metabolism Fluorescence High-Throughput Screening Assays - methods Levodopa - biosynthesis Microfluidics - methods Multidisciplinary Sciences Mutagenesis Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Science & Technology Science & Technology - Other Topics Yarrowia - genetics Yarrowia - metabolism |
| title | Sorting for secreted molecule production using a biosensor-in-microdroplet approach |
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