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Questions and Answers
What is a primary challenge in constructing three-dimensional DNA origami?
What is a primary challenge in constructing three-dimensional DNA origami?
What is one function of the caDNAno program in DNA origami design?
What is one function of the caDNAno program in DNA origami design?
What characteristic shapes are mentioned as part of DNA origami structures used in drug delivery?
What characteristic shapes are mentioned as part of DNA origami structures used in drug delivery?
How is doxorubicin typically utilized in the context of DNA origami?
How is doxorubicin typically utilized in the context of DNA origami?
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In a subcutaneous breast tumor model, which DNA origami shape demonstrated optimal accumulation at tumor regions?
In a subcutaneous breast tumor model, which DNA origami shape demonstrated optimal accumulation at tumor regions?
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What is a potential benefit of using DNA origami as a drug delivery system in cancer therapy?
What is a potential benefit of using DNA origami as a drug delivery system in cancer therapy?
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What imaging technique is mentioned for analyzing the structure of DNA origami?
What imaging technique is mentioned for analyzing the structure of DNA origami?
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What role does the 'Slice panel' feature in caDNAno serve?
What role does the 'Slice panel' feature in caDNAno serve?
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What was observed regarding the accumulation of Doxorubicin at tumor sites?
What was observed regarding the accumulation of Doxorubicin at tumor sites?
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What is a key benefit of using DNA-based nanostructures in therapeutics?
What is a key benefit of using DNA-based nanostructures in therapeutics?
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What potential advantage does DOX/origami have during in vivo delivery?
What potential advantage does DOX/origami have during in vivo delivery?
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Which of the following is NOT an application of DNA structures mentioned?
Which of the following is NOT an application of DNA structures mentioned?
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In what way do DNA nanostructures impact advanced health sciences?
In what way do DNA nanostructures impact advanced health sciences?
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What aspect of DNA origami allows for straightforward characterization?
What aspect of DNA origami allows for straightforward characterization?
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How do DNA origami structures contribute to cancer treatment?
How do DNA origami structures contribute to cancer treatment?
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What was the fluorescence signal like for free QDs and QD-M13 DNA at tumor sites?
What was the fluorescence signal like for free QDs and QD-M13 DNA at tumor sites?
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What aspect of DNA origami construction is represented in the initial phase of the design process?
What aspect of DNA origami construction is represented in the initial phase of the design process?
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Why are staple strands important in the DNA origami method?
Why are staple strands important in the DNA origami method?
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What is the typical length of the scaffold strand used in DNA origami?
What is the typical length of the scaffold strand used in DNA origami?
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What is the main goal of refining the helical domain lengths during the DNA origami design?
What is the main goal of refining the helical domain lengths during the DNA origami design?
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Which phase involves the design of complementary sequences for staple strands?
Which phase involves the design of complementary sequences for staple strands?
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What does the folding path represent in the design of DNA origami?
What does the folding path represent in the design of DNA origami?
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At which phase can staple strands be adjusted to improve the overall structure?
At which phase can staple strands be adjusted to improve the overall structure?
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What is one of the main functions of breaking and merging staple strands after refinement?
What is one of the main functions of breaking and merging staple strands after refinement?
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Study Notes
Introduction to DNA origami
- The field of structural DNA nanotechnology began around 30 years ago with Ned Seeman's pioneering research on DNA junctions and lattices.
- The invention of DNA origami in 2006 was pivotal in the rapid development of this field.
- DNA origami involves folding a long single-stranded "DNA scaffold strand" into a custom shape using short synthetic strands known as "staples."
Design of Scaffolded DNA Origami
- This involves a five-phase process, including manual and automatic design steps:
- Generation of a block diagram: This three-dimensional diagram represents the design with the scaffold as cylinders along the helical axis.
- Generation of a folding path: This sequence of moves guides the scaffold strand towards the final structure.
- Design of staple strands: These short DNA strands bind to the scaffold strand at specific locations to hold it in place.
- Refinement of helical domain lengths: This step involves adjusting crossovers to minimize strain and ensuring a better structure.
- Breaking and merging of strands: This final step can improve the stability or flexibility of the structure.
One-pot reaction
- Involves combining ~200-250 staple and scaffold strands.
- Annealed from 95°C to 20°C in a PCR machine over 2 hours.
- Structure then visualized using Atomic Force Microscopy (AFM).
Design of Three-dimensional DNA Origami
- The first three-dimensional origami was designed in 2009, taking the form of a cube.
- The complexity of this design lies in creating staple strands that hold the structure together at specific angles.
caDNAno
- This program facilitates DNA origami designing.
- Its features include:
- Slice panel: Helps visualize x-y cross sections of the honeycomb lattice.
- Path panel: Allows for detailed editing at the nucleotide level.
- Render model: Provides a 3D model for visualizing the shape.
- Auto-staple button: Generates a default set of staple paths.
Role of DNA Origami in Cancer Therapy
- Used as a biocompatible delivery system for drugs with potential to target tumor cells directly.
- DNA origami can encapsulate doxorubicin, a common chemotherapy drug, and deliver it to tumor cells.
- Doxorubicin, known for its cytotoxic properties, can effectively kill cancer cells.
DNA origami as an in vivo drug delivery vehicle for cancer therapy
- DNA origami structures, specifically triangle-shaped ones, exhibit optimal tumor passive targeting accumulation.
- Research involved comparing different DNA origami shapes, including M13 DNA, triangle, tube, and square.
Biodistribution in a subcutaneous breast tumor model
- Research involved using M13 DNA and various DNA origami shapes, all containing equivalent doses of quantum dots.
- Triangle DNA origami demonstrated optimal accumulation at the tumor regions compared to square and tube designs.
- Fluorescence signal was observed at the tumor sites, whereas signal strength was weaker for free QDs and QD-M13 DNA.
Drug loading and distribution of Doxorubicin within tumor tissues
- Triangle DNA origami was loaded with doxorubicin.
- Accumulation of doxorubicin (DOX) and DOX/origami was observed at the tumor sites.
- Fluorescence showed higher doxorubicin concentration at the tumor sites in the DOX/origami-treated group.
- Drug delivery by DNA origami carriers was observed primarily surrounding blood vessels of tumor regions.
Conclusion
- DNA origami's ability to create nano-objects with predetermined functions and their sophisticated implementations progressed quickly.
- The advantages of DNA-based nanostructures in therapeutics include their biocompatibility and biodegradability.
- Their unique properties facilitate straightforward characterization and engineering for targeting and drug delivery.
Other Applications
- Delivery of small interfering RNA (siRNA).
- CpG triggered immunostimulation.
- Efficient cellular delivery with rectangular DNA origamis coated with virus capsid proteins.
- Virus-inspired membrane-encapsulated spherical DNA origami vehicles for reduced immune activation and enhanced bioavailability.
- DNA origami that can form ion channels in lipid membranes.
- Structures that can be inserted into cell membranes to create pores that kill cancer cells.
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Description
Explore the fascinating world of DNA origami and its significance in structural DNA nanotechnology. This quiz covers the foundational concepts, including the design phases and the roles of scaffold and staple strands in creating intricate DNA shapes.
