Journal
SCIENCE
Volume 351, Issue 6280, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.aad6253
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Funding
- Synthetic Genomics (SGI)
- Defense Advanced Research Projects Agency's Living Foundries program [HR0011-12-C-0063]
- NIH from National Institute of General Medical Sciences [P41GM103412]
- Fannie and John Hertz Graduate Fellowship
- Massachusetts Institute of Technology (MIT) Center for Bits and Atoms
- MIT Department of Physics
- National Institute of Standards and Technology
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We used whole-genome design and complete chemical synthesis to minimize the 1079-kilobase pair synthetic genome of Mycoplasma mycoides JCVI-syn1.0. An initial design, based on collective knowledge of molecular biology combined with limited transposon mutagenesis data, failed to produce a viable cell. Improved transposon mutagenesis methods revealed a class of quasi-essential genes that are needed for robust growth, explaining the failure of our initial design. Three cycles of design, synthesis, and testing, with retention of quasi-essential genes, produced JCVI-syn3.0 (531 kilobase pairs, 473 genes), which has a genome smaller than that of any autonomously replicating cell found in nature. JCVI-syn3.0 retains almost all genes involved in the synthesis and processing of macromolecules. Unexpectedly, it also contains 149 genes with unknown biological functions. JCVI-syn3.0 is a versatile platform for investigating the core functions of life and for exploring whole-genome design.
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