Towards a Synthetic Chloroplast

Background The evolution of eukaryotic cells is widely agreed to have proceeded through a series of endosymbiotic events between larger cells and proteobacteria or cyanobacteria, leading to the formation of mitochondria or chloroplasts, respectively. Engineered endosymbiotic relationships between different species of cells are a valuable tool for synthetic biology, where engineered pathways based on two species could take advantage of the unique abilities of each mutualistic partner. Results We explored the possibility of using the photosynthetic bacterium Synechococcus elongatus PCC 7942 as a platform for studying evolutionary dynamics and for designing two-species synthetic biological systems. We observed that the cyanobacteria were relatively harmless to eukaryotic host cells compared to Escherichia coli when injected into the embryos of zebrafish, Danio rerio, or taken up by mammalian macrophages. In addition, when engineered with invasin from Yersinia pestis and listeriolysin O from Listeria monocytogenes, S. elongatus was able to invade cultured mammalian cells and divide inside macrophages. Conclusion Our results show that it is possible to engineer photosynthetic bacteria to invade the cytoplasm of mammalian cells for further engineering and applications in synthetic biology. Engineered invasive but non-pathogenic or immunogenic photosynthetic bacteria have great potential as synthetic biological devices.

[1]  P. Silver,et al.  Emergent cooperation in microbial metabolism , 2010, Molecular systems biology.

[2]  S. Basu,et al.  A synthetic multicellular system for programmed pattern formation , 2005, Nature.

[3]  A. Douglas,et al.  Photosynthetic symbioses in animals. , 2008, Journal of experimental botany.

[4]  Priscilla E. M. Purnick,et al.  The second wave of synthetic biology: from modules to systems , 2009, Nature Reviews Molecular Cell Biology.

[5]  Marcus Taupp,et al.  Growth, Virulence, and Immunogenicity of Listeria monocytogenes aro Mutants , 2004, Infection and Immunity.

[6]  J. Sadoff,et al.  Attenuated Shigella as a DNA Delivery Vehicle for DNA-Mediated Immunization , 1995, Science.

[7]  P. Silver,et al.  Engineering Cyanobacteria To Synthesize and Export Hydrophilic Products , 2010, Applied and Environmental Microbiology.

[8]  A. Douglas,et al.  Nutritional interactions in insect-microbial symbioses: aphids and their symbiotic bacteria Buchnera. , 1998, Annual review of entomology.

[9]  K. Jeon Change of Cellular “Pathogens” into Required Cell Components a , 1987, Annals of the New York Academy of Sciences.

[10]  J. Cirillo Exploring a novel perspective on pathogenic relationships. , 1999, Trends in microbiology.

[11]  Christopher A. Voigt,et al.  Environmentally controlled invasion of cancer cells by engineered bacteria. , 2006, Journal of molecular biology.

[12]  F. Arnold,et al.  Engineering microbial consortia: a new frontier in synthetic biology. , 2008, Trends in biotechnology.

[13]  Wenying Shou,et al.  Synthetic cooperation in engineered yeast populations , 2007, Proceedings of the National Academy of Sciences.

[14]  R. Buchsbaum Chick Tissue Cells and Chlorella in Mixed Cultures , 1937, Physiological Zoology.

[15]  P. Silver,et al.  Dynamics in the mixed microbial concourse. , 2010, Genes & development.

[16]  J. Chin,et al.  Modular approaches to expanding the functions of living matter , 2006, Nature chemical biology.

[17]  B. Lang,et al.  Mitochondrial Evolution , 1999 .

[18]  P. Youngman,et al.  Bacillus subtilis expressing a haemolysin gene from Listeria monocytogenes can grow in mammalian cells , 1990, Nature.

[19]  Shuanglin Xiang,et al.  Short hairpin RNA–expressing bacteria elicit RNA interference in mammals , 2006, Nature Biotechnology.

[20]  Pamela A. Silver,et al.  Engineering a Synthetic Dual-Organism System for Hydrogen Production , 2009, Applied and Environmental Microbiology.

[21]  B. Hall,et al.  Intracellular invasion of green algae in a salamander host , 2011, Proceedings of the National Academy of Sciences.

[22]  J. Banfield,et al.  Community structure and metabolism through reconstruction of microbial genomes from the environment , 2004, Nature.

[23]  J. Hasty,et al.  Synchronizing genetic relaxation oscillators by intercell signaling , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[24]  P. Sansonetti,et al.  Life on the inside: the intracellular lifestyle of cytosolic bacteria , 2009, Nature Reviews Microbiology.

[25]  A. van Oudenaarden,et al.  Snowdrift game dynamics and facultative cheating in yeast , 2009, Nature.

[26]  M. Parniske,et al.  Evolution of root endosymbiosis with bacteria: How novel are nodules? , 2009, Trends in plant science.

[27]  B. Lang,et al.  Mitochondrial evolution. , 1999, Science.

[28]  T. Meyer,et al.  Host-microbe interactions: bacteria. , 2008, Current opinion in microbiology.

[29]  Pascale Fung,et al.  Listeriolysin O is essential for virulence of Listeria monocytogenes: direct evidence obtained by gene complementation , 1989, Infection and immunity.

[30]  E. Muñoz-Elías,et al.  Carbon metabolism of intracellular bacteria , 2006, Cellular microbiology.

[31]  P. Lockhart,et al.  The origin of plastids , 2008, Philosophical Transactions of the Royal Society B: Biological Sciences.

[32]  Dennis L. Taylor,et al.  Artificially Induced Symbiosis Between Marine Flagellates and Vertebrate Tissues in Culture , 1978 .

[33]  H. Bouwer,et al.  Intracytoplasmic growth and virulence of Listeria monocytogenes auxotrophic mutants , 1993, Infection and immunity.

[34]  D. Bermudes,et al.  Live bacteria as anticancer agents and tumor-selective protein delivery vectors. , 2002, Current opinion in drug discovery & development.

[35]  W. Goebel,et al.  Microinjection and growth of bacteria in the cytosol of mammalian host cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[36]  K. Timmis,et al.  Towards elucidation of microbial community metabolic pathways: unravelling the network of carbon sharing in a pollutant-degrading bacterial consortium by immunocapture and isotopic ratio mass spectrometry. , 1999, Environmental microbiology.

[37]  N. Forbes Engineering the perfect (bacterial) cancer therapy , 2010, Nature Reviews Cancer.

[38]  R. Buchsbaum,et al.  AN ARTIFICIAL SYMBIOSIS. , 1934, Science.

[39]  L Margulis,et al.  Words as battle cries--symbiogenesis and the new field of endocytobiology. , 1990, Bioscience.

[40]  W. Goebel,et al.  Hpt, a bacterial homolog of the microsomal glucose- 6-phosphate translocase, mediates rapid intracellular proliferation in Listeria , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[41]  Christopher A. Voigt,et al.  Synthesis of methyl halides from biomass using engineered microbes. , 2009, Journal of the American Chemical Society.

[42]  M. Rumpho,et al.  Solar-powered sea slugs. Mollusc/algal chloroplast symbiosis. , 2000, Plant physiology.

[43]  G. Tattersall,et al.  Embryonic motility and hatching success of Ambystoma maculatum are influenced by a symbiotic alga , 2008 .

[44]  Samuel I. Miller,et al.  Lipid A mutant Salmonella with suppressed virulence and TNFα induction retain tumor-targeting in vivo , 1999, Nature Biotechnology.

[45]  S. Falkow,et al.  Identification of invasin: A protein that allows enteric bacteria to penetrate cultured mammalian cells , 1987, Cell.

[46]  T. Kleine,et al.  DNA transfer from organelles to the nucleus: the idiosyncratic genetics of endosymbiosis. , 2009, Annual review of plant biology.