A Pressure Test to Make 10 Molecules in 90 Days: External Evaluation of Methods to Engineer Biology.

Autor:	Casini A; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Broad Institute of MIT and Harvard , Cambridge , Massachusetts 02142 , United States., Chang FY; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Eluere R; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Broad Institute of MIT and Harvard , Cambridge , Massachusetts 02142 , United States., King AM; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Young EM; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Dudley QM; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Department of Chemical and Biological Engineering, Center for Synthetic Biology , Northwestern University , Evanston , Illinois 60208 , United States., Karim A; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Department of Chemical and Biological Engineering, Center for Synthetic Biology , Northwestern University , Evanston , Illinois 60208 , United States., Pratt K; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Broad Institute of MIT and Harvard , Cambridge , Massachusetts 02142 , United States., Bristol C; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Broad Institute of MIT and Harvard , Cambridge , Massachusetts 02142 , United States., Forget A; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Broad Institute of MIT and Harvard , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Ghodasara A; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Warden-Rothman R; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Gan R; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Department of Chemical and Biological Engineering, Center for Synthetic Biology , Northwestern University , Evanston , Illinois 60208 , United States., Cristofaro A; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Borujeni AE; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Ryu MH; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Li J; Department of Chemical and Biological Engineering, Center for Synthetic Biology , Northwestern University , Evanston , Illinois 60208 , United States., Kwon YC; Department of Chemical and Biological Engineering, Center for Synthetic Biology , Northwestern University , Evanston , Illinois 60208 , United States., Wang H; Department of Chemical and Biological Engineering, Center for Synthetic Biology , Northwestern University , Evanston , Illinois 60208 , United States., Tatsis E; Department of Biological Chemistry , John Innes Centre , Norwich NR4 7UH , United Kingdom., Rodriguez-Lopez C; Department of Biological Chemistry , John Innes Centre , Norwich NR4 7UH , United Kingdom., O'Connor S; Department of Biological Chemistry , John Innes Centre , Norwich NR4 7UH , United Kingdom., Medema MH; Bioinformatics Group , Wageningen University , Wageningen 6708 PB , The Netherlands., Fischbach MA; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Department of Bioengineering and Chemistry, Engineering & Medicine for Human Health , Stanford University , Stanford , California 94305 , United States., Jewett MC; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Department of Chemical and Biological Engineering, Center for Synthetic Biology , Northwestern University , Evanston , Illinois 60208 , United States., Voigt C; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Broad Institute of MIT and Harvard , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States., Gordon DB; The Foundry , 75 Ames Street , Cambridge , Massachusetts 02142 , United States.; Broad Institute of MIT and Harvard , Cambridge , Massachusetts 02142 , United States.; Synthetic Biology Center, Department of Biological Engineering , Massachusetts Institute of Technology , Cambridge , Massachusetts 02139 , United States.
Jazyk:	angličtina
Zdroj:	Journal of the American Chemical Society [J Am Chem Soc] 2018 Mar 28; Vol. 140 (12), pp. 4302-4316. Date of Electronic Publication: 2018 Mar 16.
DOI:	10.1021/jacs.7b13292
Abstrakt:	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.
Databáze:	MEDLINE
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