Developing Genome Transplantation for Non-mycoplasma Bacteria to Enhance the Creation of Synthetic Cells

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In 2010, researchers constructed a synthetic bacterial cell whose genome was synthesized from laboratory chemicals. It was anticipated that this first synthetic cell would be followed by many others constructed by scientists all over the world. Indeed, researchers anticipated that bacteria of all types would be designed and constructed to solve human problems in medicine, energy, industry, environmental remediation and basic research. That has yet to happen because it is difficult to transplant a rationally designed synthetic genome into a suitable recipient bacterial cell. In the project proposed here, the original inventors of the genome transplantation procedure will adapt the technique so that it can be applied to other bacteria. This will allow academic and industrial bacteriologists and synthetic biologists to build new bacteria with remarkable capabilities. In addition to removing a huge technical hurdle stalling the construction of synthetic cells, this project will facilitate the training of the next generation of scientists in the field of synthetic biology and will be used as a platform for discussions with the scientific community, industry and public. Genome Transplantation, the insertion of a complete bacterial genome into a suitable recipient cell so that the donated genome commandeers the recipient cell to produce a new cell with the genotype and phenotype of the donated genome, was the fundamental technical development that enabled the construction of the first bacterium with a synthetic genome. At the announcement of the creation of the world’s first synthetic cell, JCVI-syn1.0, the work was heralded as a major accomplishment for humans. To date, genome transplantation has only been achieved using a small subset of the atypical bacteria called mycoplasmas. The goals of this project are to investigate the mechanism of genome transplantation, develop genome transplantation for at least two species of non-mycoplasma bacteria, and use microfluidics technology to automate and improve the efficiency of genome transplantation. A potential outcome is to establish a set of rules that synthetic cell builders can use to transplant isolated whole synthetic bacterial genomes and produce new synthetic cells with desired properties.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.