Team:Berkeley

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[https://2010.igem.org/Team:Berkeley/Project_Overview Project Overview]
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[https://2010.igem.org/Team:Berkeley/Choanoflagellates Choanoflagellates]  
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[https://2010.igem.org/Team:Berkeley/Parts Parts]  
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[https://2010.igem.org/Team:Berkeley/Clotho Clotho]
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[https://2010.igem.org/Team:Berkeley/Human_Practices Human Practices]
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[https://2010.igem.org/Team:Berkeley/Notebooks Notebooks]
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<big><font size="5" face="Comic Sans"> Abstract</font> </big> <br>
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The ability to manipulate the DNA of an organism is vital to many modern fields of biology. While we have perfected this in common research species such as E. coli, yeast and mouse cells, it is still impossible to transform many other species researchers study. Our project is an attempt to develop transgenics ) techniques for a family of single celled organisms called choanoflagellates. These species are interesting to researchers because they are the closest living relative to our microbial ancestor that became the first multicellular animal. Nicole King, here at UC Berkeley, and other researchers across the globe who study these little creatures are hindered by the inability to genetically manipulate them.
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The Berkeley iGEM 2010 team is applying synthetic biology to this problem. We are engineering bacteria that can deliver DNA into the choano. Choanos are predatory, which makes our job a bit simpler. Once our bacteria is engulfed by the choano, it is programmed to burst using a self-lysis device. Proteins we have placed inside the bacteria will then go into action. First, we have designed a vacuole-buster device that will burst the small food membrane holding the bacteria inside the choano, spewing the contents into the cytoplasm of the cell. In the cytoplasm, a transposon/transposase device tagged with a nuclear localization device will move to the nucleus. In the nucleus, the transposase will splice the transposon into the choanoflagellate DNA.
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<font size="5" face="Comic Sans">Acknowledgements</font> <br>
 
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We thank our wonderful advisors: Chris Anderson, Terry Johnson, and Tim Hsiau for their guidance and support. We also thank our generous sponsors:<br>
 
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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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<a href="http://www.synberc.org/">
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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.