"

From 2010.igem.org

---

Our project focused on the delivery of payloads to the cytoplasms of choanoflagellates, but its simple design makes it applicable to a host of other eukaryotes. Our design is only limited in three main aspects as to its potential targets:

• The target organism must be able to naturally phagocytose Escheria Coli.
• The target organism's phagocytic vesicles must contain cholesterols susceptible to our Vesicle-Buster device.
• The target organism must be able to survive in media that is compatible with our Self-Lysis device.

We have started looking at other organisms to deliver payloads to using our payload delivery device. Having screened about a dozen single-celled, freshwater eukaryotes for phagocytosis of E. coli and ability to survive in various concentrations of LB, we are now focusing our efforts on [http://en.wikipedia.org/wiki/Tetrahymena Tetrahymena], [http://en.wikipedia.org/wiki/Blepharisma Blepharisma] and [http://en.wikipedia.org/wiki/Paramecium Paramecia]. We have already determined that these three types of organisms will readily eat E. coli and are capable of surviving in mixtures of their culturing media and LB. The images below show bacteria expressing GFP being ingested by these three organisms. The next step is to include the payload delivery device and start assaying them.

Tetrahymena	Tetrahymena: Phase contrast	Tetrahymena: Phase contrast and GFP	Tetrahymena: GFP only
Blepharisma	Blepharisma: Phase contrast	Blepharisma: Phase contrast and GFP	Blepharisma: GFP only
Paramecia	Paramecia: Phase contrast	Paramecia: Phase contrast and GFP	Paramecia: GFP only