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  A pressure test to make 10 molecules in 90 days: external evaluation of methods to engineer biology

Casini, A., Chang, F.-Y., Eluere, R., King, A. M., Young, E. M., Dudley, Q. M., et al. (2018). A pressure test to make 10 molecules in 90 days: external evaluation of methods to engineer biology. Journal of the American Chemical Society, 140(12), 4302-4316. doi:10.1021/jacs.7b13292.

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 Creators:
Casini, Arturo, Author
Chang, Fang-Yuan, Author
Eluere, Raissa, Author
King, Andrew M., Author
Young, Eric M., Author
Dudley, Quentin M., Author
Karim, Ashty, Author
Pratt, Katelin, Author
Bristol, Cassandra, Author
Forget, Anthony, Author
Ghodasara, Amar, Author
Warden-Rothman, Robert, Author
Gan, Rui, Author
Cristofaro, Alexander, Author
Borujeni, Amin Espah, Author
Ryu, Min-Hyung, Author
Li, Jian, Author
Kwon, Yong-Chan, Author
Wang, He, Author
Tatsis, Evangelos, Author
Rodriguez-Lopez, Carlos, AuthorO'Connor, Sarah E.1, Author           Medema, Marnix H., AuthorFischbach, Michael A., AuthorJewett, Michael C., AuthorVoigt, Christopher, AuthorGordon, D. Benjamin, Author more..
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1External Organizations, ou_persistent22              

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 Abstract: 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 cellfree 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 albidof lavus, Streptomyces coelicolor, and Streptomyces albovinaceus), established two cell-free systems, and performed 690 assays developed in-house for the molecules.

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 Dates: 2018-02-262018
 Publication Status: Published in print
 Pages: -
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 Rev. Type: -
 Identifiers: Other: SOC081
DOI: 10.1021/jacs.7b13292
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Title: Journal of the American Chemical Society
  Other : J. Am. Chem. Soc.
  Abbreviation : JACS
Source Genre: Journal
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Publ. Info: Washington, DC : American Chemical Society
Pages: - Volume / Issue: 140 (12) Sequence Number: - Start / End Page: 4302 - 4316 Identifier: ISSN: 0002-7863
CoNE: https://pure.mpg.de/cone/journals/resource/954925376870