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Triggered Sequestration with DNA Nanostructures: A New Drug Delivery Method August 14, 2006 iGEM Week 10: Progress Report Tiffany Chan, Katherine Fifer,

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Presentation on theme: "Triggered Sequestration with DNA Nanostructures: A New Drug Delivery Method August 14, 2006 iGEM Week 10: Progress Report Tiffany Chan, Katherine Fifer,"— Presentation transcript:

1 Triggered Sequestration with DNA Nanostructures: A New Drug Delivery Method August 14, 2006 iGEM Week 10: Progress Report Tiffany Chan, Katherine Fifer, Valerie Lau, Matthew Meisel

2 Overview Project goal Last week’s progress test AscI digest Microcon filtration [Mg 2+ ] and [oligo] titration This week’s goals

3 Project goal To create a scaffolded DNA nanostructure that protects a ligand from a degradation enzyme When the ligand is presented on the outside of the nanostructure, we expect there to be no protection

4 AscI digest Two ss DNAs (the “attachment DNA” and the “oligo ligand”) with 15-bp complimentary stretch were briefly incubated at room temperature The double-stranded construct was then treated with AscI under four different digestion conditions 25 kb+ 10 kb+ 25 kb+ milliunits AscI: digestion time: (minutes)

5 Microcon filtration Samples are pipetted into the Microcon filter and are successively diluted and centrifuged Results: relatively good oligo removal, but poor nanostructure yields 1 kb+unpurifiedsuccessive flow-throughs scaffoldretentate Image courtesy Millipore

6 More PEG experiments PEG titration performed for each set of folding conditions pelletpelletetc…supern.

7 Other filtration options Microcon use more material—is yield loss constant? use a detergent to loosen nanostructures from membrane (Triton X-100?) Sepharose spin columns large molecules elute first homemade: Datta et al. (2003), Anal. Biochem., 317, p Clonetech Prespin Load sample Centrifuge (large molecules elute first) Image courtesy Clonetech

8 This week’s goals Continue filtration/purification trials AscI digest of folded nanostructure


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