Goals & Success Criteria
Our Criteria for Success
DNA Origami Box Design
- Box is thermodynamically stable
- Box design is capable of docking anti-CD3 antibody
- Box has a viable lock and key mechanism
- Anti-CD3 antibody successfully attaches inside of box
Evaluation of Target Specificity to Prostate Cancer Cells
- Anti-CD3 antibody binds to T cells
- Box + anti-CD3 antibody binds to T cells
- Design RNA aptamer that binds to PSMA receptor (aptamer 1)
- Box + aptamer 1 binds to PSMA receptor
Validation of Function Upon Binding to Prostate Cancer Cells
- T cells are recruited by box + anti-CD3 antibody + aptamer
- T cells are activated by box + anti-CD3 antibody + aptamer
- Box + anti-CD3 antibody + aptamer 1 opens upon binding to PSMA receptor
- Box + anti-CD3 antibody + aptamers 1 and 2 opens upon binding to both PSMA and NTSR1 receptors
- In vitro testing using LNCaP cells
Objectives
The objectives for this project are as follows:
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Designing and synthesizing an appropriate DNA Origami Box via DNA/RNA nanostructure tools, and validating the formation of our DNA origami box using gel electrophoresis and TEM
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Attaching the anti-CD3 antibody to the box via antibody docking, and validating the attachment of the anti-CD3 antibody by gel electrophoresis and TEM
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Verifying that our chosen aptamer binds to PSMA with ELISA
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Confirming that our chosen anti-CD3 antibody binds to T-cells using flow cytometry
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Verifying that the box will bind to prostate cancer cells and open via flow cytometry
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Checking for T-cell activation and recruitment to treat prostate cancer by flow cytometry and a transwell migration assay