Download Designing DNA Nanostructures to encapsulate and

Designing DNA Nanostructures to
encapsulate and release proteins
iGEM
August 21, 2006
Katherine Fifer
with Valerie Lau, Matthew Meisel, and Tiffany Chan
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).
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
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
Motivation: Why DNA?

The power of DNA




Watson-Crick base pairing is enormously
strong
Self-assembly
Highly programmable, designable
Specificity
Scaffolded Oragami
scaffold
staples
Scaffolded Oragami
Progress



Built a number of barrel designs
Exciting EM Images
Purifying Nanostructures (nearly there
after 1 month of trials)
Exciting EM Images
Exciting EM Images
EM Images (snakes on a grid)


Images courtesy Shawn Douglas
c5.0 barrel
(10 nM), 0.7%
uranyl
formate
Appear to be
lining up end
to end,
probably
because of
the stain
EM Images
Images courtesy Shawn Douglas
To be continued

Protection assays



Can a protein be
protected from
protease if attached
inside the box
Lid attachment
Lid removal

Protease or enzyme
Strand displacement
Protease or enzyme
Protein or
DNA ligand
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