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Hutchison, Clyde A.; Chuang, Ray-Yuan; Noskov, Vladimir N.; Assad-Garcia, Nacyra; Deerinck, Thomas J.; Ellisman, Mark H.; Gill, John; Kannan, Krishna; Karas, Bogumil J.; Ma, Li; Pelletier, James F.; Qi, Zhi-Qing; Richter, R. Alexander; Strychalski, Elizabeth A.; Sun, Lijie; Suzuki, Yo; Tsvetanova, Billyana; Wise, Kim S.; Smith, Hamilton O.; Glass, John I.; Merryman, Chuck; Gibson, Daniel G.; Venter, J. Craig
Science, 03/2016, Volume: 351, Issue: 6280Journal Article
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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