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Direct transfer of whole genomes from bacteria to yeast

Abstract

Transfer of genomes into yeast facilitates genome engineering for genetically intractable organisms, but this process has been hampered by the need for cumbersome isolation of intact genomes before transfer. Here we demonstrate direct cell-to-cell transfer of bacterial genomes as large as 1.8 megabases (Mb) into yeast under conditions that promote cell fusion. Moreover, we discovered that removal of restriction endonucleases from donor bacteria resulted in the enhancement of genome transfer.

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Figure 1: Effects of disrupting restriction-modification systems on genome transfer.
Figure 2: Modes of cell-to-cell genome transfer from bacterium to yeast.

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Acknowledgements

We thank J. Firstenhaupt for preparing Figure 2a. This work was supported by Synthetic Genomics, Inc. B.J.K. was supported by the National Science and Engineering Research Council of Canada Postdoctoral Fellowships Program and by Synthetic Genomics, Inc. Y.S. was supported by the US Defense Advanced Research Projects Agency contract N66001-12-C-4039.

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Authors

Contributions

B.J.K., J.J., J.C.V., P.D.W., D.G.G., C.A.H., H.O.S. and Y.S. designed the research. B.J.K., J.J., L.S., L.M., J.S., A.R., P.D.W., D.G.G., C.A.H. and Y.S. performed experiments. G.M.G., M.J.M. and E.A.W. performed genome sequencing and analysis. B.J.K., J.J., J.S., A.R., P.D.W., D.G.G., C.A.H., H.O.S. and Y.S. wrote the paper.

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Correspondence to Bogumil J Karas or Yo Suzuki.

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Competing interests

J.C.V. is chief executive officer and co-chief scientific officer, H.O.S. is co-chief scientific officer and a member of the board of directors, C.A.H. is chairman of the scientific advisory board, and D.G.G. is a vice president of Synthetic Genomics, Inc. All four of these authors and the J. Craig Venter Institute hold shares of this company.

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Supplementary Figures 1–6, Supplementary Tables 1 and 2 and Supplementary Notes 1–7 (PDF 663 kb)

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Karas, B., Jablanovic, J., Sun, L. et al. Direct transfer of whole genomes from bacteria to yeast. Nat Methods 10, 410–412 (2013). https://doi.org/10.1038/nmeth.2433

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