A Pressure Test to Make 10 Molecules in 90 Days: External Evaluation of Methods to Engineer Biology
Name
Voigt_A Pressure Test to Make 10 Molecules in 90 Days.pdf
Description
Accepted version
Size
5.45 MB
Format
Unknown
Checksum (MD5)
8a301c70b86e0e329fbb0571b0bd9859
Author(s)
Casini, Arturo
Chang, Fang-Yuan
Eluere, Raissa
King, Andrew M.
Young, Eric M.
Dudley, Quentin M.
Karim, Ashty
Pratt, Katelin
Bristol, Cassandra
Forget, Anthony
Date Issued
February 2018
Journal
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Version
Author's final manuscript
Abstract
© 2018 American Chemical Society. Centralized facilities for genetic engineering, or "biofoundries", offer the potential to design organisms to address emerging needs in medicine, agriculture, industry, and defense. The field has seen rapid advances in technology, but it is difficult to gauge current capabilities or identify gaps across projects. To this end, our foundry was assessed via a timed "pressure test", in which 3 months were given to build organisms to produce 10 molecules unknown to us in advance. By applying a diversity of new approaches, we produced the desired molecule or a closely related one for six out of 10 targets during the performance period and made advances toward production of the others as well. Specifically, we increased the titers of 1-hexadecanol, pyrrolnitrin, and pacidamycin D, found novel routes to the enediyne warhead underlying powerful antimicrobials, established a cell-free system for monoterpene production, produced an intermediate toward vincristine biosynthesis, and encoded 7802 individually retrievable pathways to 540 bisindoles in a DNA pool. Pathways to tetrahydrofuran and barbamide were designed and constructed, but toxicity or analytical tools inhibited further progress. In sum, we constructed 1.2 Mb DNA, built 215 strains spanning five species (Saccharomyces cerevisiae, Escherichia coli, Streptomyces albidoflavus, Streptomyces coelicolor, and Streptomyces albovinaceus), established two cell-free systems, and performed 690 assays developed in-house for the molecules.
MIT Department
Massachusetts Institute of Technology. Synthetic Biology Center
Massachusetts Institute of Technology. Department of Biological Engineering
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1021/jacs.7b13292
