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Designing DNA Nanostructures to encapsulate and release proteins iGEM August 21, 2006 Katherine Fifer with Valerie Lau, Matthew Meisel, and Tiffany Chan
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Background: What is iGEM? iGEM - intercollegiate Genetically Engineered Machines Competition between about 40 schools in November Emphasis on biobricking parts to enable building of genetic circuits Biobricking parts makes them interchangeable (usually).
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DNA Nanostructures Overview Can stick double helices together A few basic shapes have already been made A 1.7-kilobase single-stranded DNA that folds into a nanoscale octahedron WILLIAM M. SHIH, JOEL D. QUISPE & GERALD F. JOYCE Nature 427, 618 ミ 621 (2004); doi:10.1038/nature02307
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Motivation: For what purpose? Creating microenvironments Encapsulating, releasing proteins Specificity - arriving at certain targets Timed release A new drug delivery model? Thrombin Model Proof of principle
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Motivation: Why DNA? The power of DNA Watson-Crick base pairing is enormously strong Self-assembly Highly programmable, designable Specificity
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Scaffolded Oragami scaffold staples
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Scaffolded Oragami
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Progress Built a number of barrel designs Exciting EM Images Purifying Nanostructures (nearly there after 1 month of trials)
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Exciting EM Images
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EM Images (snakes on a grid) c5.0 barrel (10 nM), 0.7% uranyl formate Appear to be lining up end to end, probably because of the stain Images courtesy Shawn Douglas
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EM Images Images courtesy Shawn Douglas
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To be continued Protection assays Can a protein be protected from protease if attached inside the box Lid attachment Lid removal Strand displacement Protease or enzyme Protein or DNA ligand
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Acknowledgements iGEM team - Valerie Lau, Matthew Meisel, Tiffany Chan, Perry Tsai, Lewis Hahn, Jeff Lau, Hetmann Hsieh, Zhipeng Sun, David Ramos iGEM TFs - Shawn Douglas, Nick Stroustrup, Chris Doucette iGEM advisers - Dr. William Shih, Dr. George Church, Dr. Pamela Silver, Dr. Alain Viel, Dr. Jagesh Shah, Dr. Radhika Nagpal PRISE Shawn Douglas Dr. William Shih
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