Berkeley UC

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The necessity of cheap, available, disease free, and universally usable blood substitutes is undisputed. There are currently no blood substitutes approved for use in the US or the UK, and whole blood is almost always in short supply. Developing countries have the greatest need for blood transfusions, however many lack the necessary donation and storage infrastructure and the required number of healthy donors. To address this problem, we are developing an innovative and inexpensive blood substitute constructed from E. coli bacteria engineered to include the critical capabilities of human erythrocytes. Our bacterial system includes the ability to safely exist in the bloodstream, carry oxygen with hemoglobin, and be stored for prolonged periods in a freeze-dried state.

Support for Berkeley iGEM 2007 was generously provided by SynBERC and The Camille and Henry Dreyfus Foundation, Inc.

Project Modules
	Oxygen Carrying Our system is designed to produce Hemoglobin, Heme, and the necessary chaperones and detoxifying agents to promote the transport of oxygen throughout the bloodstream. We also investigated alternates to hemoglobin and other strategies for its production.
	The Chassis Our bacterial chassis has been heavily modified to remove its sepsis-inducing toxicity, as well as promote its ability to last longer in the bloodstream by masking it from the immune system. We performed this by lipopolysaccharide modifications as well as adding the K-capsule and O-antigen.
	The Controller The Controller is an integrated genetic circuit, comprised of two plasmids, that directs the initiation and production of the primary systems proteins through its component operons. It is composed of a pSC101 controller plasmid and an amplifiable BAC utilizing the T7 RNAP expression system.
	Genetic Self-Destruct To prevent chance of infection or unwanted proliferation after hemoglobin production, we have engineered a genetic self-destruct mechanism whereby when induced, the bacterial cell will express a genetic material-degrading toxin which kills the cell, but leaves it physically intact.
	Freeze Drying In order to enable preservation of our bacteria for long time periods, we have included the ability to produce compounds, such as hydroxyectoine, that will enable our bacteria to survive freeze-drying intact. This will dramatically increase shelf-life and decrease transport costs.