NOV 03, 2016 10:30 AM PDT

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

Speaker
  • Assistant Professor, Leslie Dan Faculty of Pharmacy, University of Toronto
    Biography
      Dr. Pardee is an Assistant professor in the Leslie Dan Faculty of Pharmacy, at the University of Toronto. He holds an Honours Bachelor of Science degree in Biological Sciences from the University of Alberta, a Master of Science degree in Natural Products Chemistry from the University of British Columbia, and a Doctor of Philosophy degree in Molecular Genetics from the University of Toronto. Following the completion of his doctoral studies, Dr. Pardee completed a Postdoctoral Fellowship under the supervision of Professor James J. Collins at the Wyss Institute at Harvard University.

      Dr. Pardee's work at Harvard combined in vitro synthetic biology and biochemical systems with materials science to build paper-based synthetic gene networks. With this system he created a method to embed freeze-dried synthetic gene networks and their complementary cellular machinery into paper. These systems remain stable without refrigeration for more than a year, and are activated by adding water. These devices provide a new venue for synthetic biologists to operate in, and a much-needed path for the safe deployment of engineered gene circuits beyond the lab. With this technology he has developed diagnostics for the Zika virus (http://www.cell.com/fulltext/S0092-8674(16)30505-0) and as well as developed a new portable manufacturing platform for therapeutics (http://www.cell.com/fulltext/S0092-8674(16)31246-6).

      Dr. Pardee's research has been published in Cell, Nature, PLoS Biology, and Genes and Development, among others.

    Abstract

    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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