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(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.



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.