Team:Berkeley/Project Overview

From 2010.igem.org

Choa's Choa's Delivery Service!

The ability to manipulate the DNA of an organism is vital to many modern fields of biology. Although we have perfected this in common research species such as E. coli, yeast and mammalian cells, it is still impossible to transform many other species. Lower metazoans in particular have several advantages over pre-existing chassis but remain out of reach because of an inability to deliver protein and DNA. From the perspective of synthetic biology, lower metazoans hold the promise of harnessing new biological pathways and developing complex biological machinery. Compared to bacteria and yeast, they are both evolutionarily closer to animals and much easier to culture. Today, however, they are generally avoided due to a lack of genetic engineering techniques. E. coli and S. cerevisia continue to dominate the field not because of their genetic or biological value, but because they were readily accessible to our predecessors. Our project was to expand synthetic biology to new lower metazoan chassis by designing a general device that can deliver protein and DNA to any phagocytic eukaryotes.

We are engineering bacteria to serve as a vector to deliver payload into choanoflagellates. Since Choanoflagellates naturally eat bacteria, our bacteria easily entered the choanoflagellate. Once our bacteria is engulfed by the choanoflagellate, it is programmed to open itself using a self-lysis device derived from the 2008 UC Berkeley iGEM team. At this point, proteins being expressed by the bacteria will be released and ready to act. A vesicle buster device will open the small food vesicle and release our payload into the cytoplasm. The payload can consist of either protein, nucleic acids, or a combination of the both. We demonstrated the functionality of the the self-lysis and vesicle buster device by delivery GFP to the cytoplasm of the choanoflagellate. Future work involves targeting protein and DNA to the nucleus in order to genetically modify the choanoflagellate. A transposon/transposase device will move to the nucleus and splice DNA in or out of the genome. Although we tested our constructs on choanoflagellates, the devices are general enough to be applied to any phagocytic organism.