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RNA-aptamers-in-droplets (RAPID) high-throughput screening for secretory phenotypes
Joseph Abatemarco1, Maen F Sarhan2,3, James M Wagner1
1Department of Chemical Engineering, The University of Texas at Austin, 200 E Dean Keeton St Stop C0400, Austin, Texas, 78712, USA.
Nature Communications
|August 25, 2017
Summary
Researchers developed RNA-aptamers-in-droplets (RAPID) to rapidly screen engineered microbes for chemical production. This ultrahigh-throughput method uses aptamers to convert product presence into fluorescence, enabling faster discovery of microbial "living foundries".
Area of Science:
- Synthetic biology
- Metabolic engineering
- Biotechnology
Background:
- Microbial engineering aims to create
- living foundries
- for high-value chemical production through iterative design-build-test cycles.
Purpose of the Study:
- To address the bottleneck in high-throughput screening of engineered microbial libraries.
- To develop a generalizable method for ultrahigh-throughput screening of microbial secreted products.
Main Methods:
- RNA-aptamers-in-droplets (RAPID) combines aptamer-based fluorescent reporters with droplet microfluidics.
- Aptamers transduce extracellular product concentration into measurable fluorescence signals.
- This enables screening of millions of microbial variants in droplets.
Main Results:
- RAPID achieved ultrahigh-throughput screening of microbial libraries.
- Tyrosine production in Saccharomyces cerevisiae was enhanced up to 28-fold.
- Recombinant protein secretion in yeast was improved up to 3-fold.
Conclusions:
- RAPID significantly expands the generality and throughput of microfluidic screening.
- This method facilitates the evolution of microbes for enhanced synthesis and secretion of valuable chemicals.
- The approach is broadly applicable for optimizing microbial cell factories.
