Rapid, Low-cost Tools for Global Health: Using Cell-free Synthetic Biology for Zika Virus Detection and the Portable Manufacture of Therapeutics.

Keith Pardee, Assistant Professor, Leslie Dan Faculty of Pharmacy, University of Toronto

22 November 2016 at 11:30

Location: BSB B136

The recent Zika virus outbreak highlights the need for low-cost diagnostics that can be rapidly developed for distribution and use in pandemic regions.  In early 2016 we developed a pipeline for the rapid design, assembly, and validation of cell-free, paper-based sensors for the detection of the Zika virus RNA genome.  This work was built upon a paper-based system that we originally published in 2014 that used freeze-dried cell-free reactions to deploy synthetic gene networks outside of the lab in a sterile and abiotic format.  By linking isothermal RNA amplification to toehold switch RNA sensors in this paper-based system, we were able to detect clinically relevant concentrations of Zika virus sequences and demonstrated specificity against closely related Dengue virus sequences.  When coupled with a novel CRISPR/Cas9-based module, our sensors were able to discriminate between viral strains with single-base resolution.  We also successfully demonstrated a simple, field-ready sample-processing workflow and detect Zika virus from the plasma of a viremic macaque.  


We have also recently extended our cell-free approach to the portable manufacture of therapeutics.  Using pellets of freeze-dried cell-free reactions we demonstrated the synthesis over 50 products.  This included the manufacture and functional validation of antimicrobial peptides, vaccines, antibody conjugates and small molecules.  Our freeze-dried biomolecular platform resolves important practical limitations to the deployment of both molecular diagnostics and protein-based therapeutics to the field and demonstrates how synthetic biology can be used to develop tools for confronting global health crises.



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