Team:Berkeley

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<big><font size="5" face="Book Antiqua"> Abstract</font> </big> <br>
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Cell surface display in ''E. coli'' tethers proteins to the outer membrane in order to localize them to the extracellular environment.  While this form of localization has allowed many novel functions to be engineered into ''E. coli'', work within this space relies on a trial and error approach rather than design principles. We worked to create a foundation of research which would make the rational design of cell surface display systems in ''E. coli''  possible.  We used a combinatorial approach to compare the ability of different display proteins to display different classes of functional proteins.  This required the development and implementation of an automated assembly method able to construct the large number of devices necessary to draw meaningful conclusions about design within this space. 
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Single-celled phagocytic eukaryotes, such as Choanoflagellates, are of great interest to developmental biologists because they may be the last living immediate precursor on the evolutionary tree to animals. These easy-to-culture and robust organisms are also a desirable low-cost eukaryotic chassis for synthetic biology, but there are few to no tools for delivering biomolecules into these organisms. We engineered E. coli to deliver proteins and/or DNA payloads into these bacteria-devouring eukaryotes. Once ingested, our E. coli are programmed to self-lyse and porate the phagosome, releasing their payloads into the cytosol. This delivery mechanism has the potential to deliver payload to any phagocytic organism with a cholesterol-based membrane. As part of our parallel software effort to rework the Clotho plugin environment and API, we made automatic biosafety handling an intrinsic feature of the core. Together, these tools provide a foundation for metazoan synthetic biology and a framework for improving safety in our field.
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===Sponsors===
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<center><font size="5" face="Book Antiqua">Our Team</font></center> <br>
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&nbsp;&nbsp;&nbsp;
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<font size="5" face="Book Antiqua">Acknowledgements</font> <br>
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We thank our wonderful advisers: Chris Anderson, Terry Johnson, and Lane Weaver for their guidance and support. We also thank our generous sponsors:<br>
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&nbsp;&nbsp;&nbsp;
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<br>&nbsp;</br>
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We would like to thank our sponsors, shown above, and our wonderful advisors Chris Anderson, Terry Johnson, Tim Hsiau, and Jin Huh for their guidance and support.

Latest revision as of 23:43, 27 October 2010

Plain header.png




ChoaChoa delivery header.png
(Pronounced like "Cocoa")

Single-celled phagocytic eukaryotes, such as Choanoflagellates, are of great interest to developmental biologists because they may be the last living immediate precursor on the evolutionary tree to animals. These easy-to-culture and robust organisms are also a desirable low-cost eukaryotic chassis for synthetic biology, but there are few to no tools for delivering biomolecules into these organisms. We engineered E. coli to deliver proteins and/or DNA payloads into these bacteria-devouring eukaryotes. Once ingested, our E. coli are programmed to self-lyse and porate the phagosome, releasing their payloads into the cytosol. This delivery mechanism has the potential to deliver payload to any phagocytic organism with a cholesterol-based membrane. As part of our parallel software effort to rework the Clotho plugin environment and API, we made automatic biosafety handling an intrinsic feature of the core. Together, these tools provide a foundation for metazoan synthetic biology and a framework for improving safety in our field.


Sponsors


               
 

We would like to thank our sponsors, shown above, and our wonderful advisors Chris Anderson, Terry Johnson, Tim Hsiau, and Jin Huh for their guidance and support.

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