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Design and synthesis of a minimal bacterial genome



Elizabeth A. Strychalski, Clyde A. Hutchinson III, Ray-Yuan Chuang, Vladimir Noskov, Nacyra Assad-Garcia, Tom J. Deerinck, Mark H. Ellisman, John Gill, Krishna Kannan, Bogumil J. Karas, Li Ma, James F. Pelletier, Zhi-Qing Qi, Alexander Richter, Lijie Sun, Yo Suzuki, Billyana Tsvetanova, Kim S. Wise, Hamilton O. Smith, John I. Glass, Chuck Merryman, Daniel G. Gibson, J. C. Venter


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.


Strychalski, E. , Hutchinson, C. , Chuang, R. , Noskov, V. , Assad-Garcia, N. , Deerinck, T. , Ellisman, M. , Gill, J. , Kannan, K. , Karas, B. , Ma, L. , Pelletier, J. , Qi, Z. , Richter, A. , Sun, L. , Suzuki, Y. , Tsvetanova, B. , Wise, K. , Smith, H. , Glass, J. , Merryman, C. , Gibson, D. and Venter, J. (2016), Design and synthesis of a minimal bacterial genome, Science, [online], science.aad6253 (Accessed April 23, 2024)
Created March 25, 2016, Updated November 10, 2018