Effects of Thermal Damage on the Formation of DNA Origami
Faculty Mentor
Ken Halvorsen, Vinod Morya
Major/Area of Research
DNA Nanotechnology (PharmD.)
Description
INTRODUCTION: In DNA nanotechnology, using DNA as a building block in fabricating nanostructures has expanded the scope of DNA-focused research. With the DNA origami method, a long single-stranded DNA scaffold is folded into nanoscale shapes using complementary staple strands of oligonucleotides. As this technique is used for many biomedical applications such as drug delivery, the structural stability of the DNA origami is essential to designing more robust and reliable nanostructures for practical use. The aim of this study is to see the effects of heat damage on the construction of these DNA origami nanostructures, using DNA triangles as a model structure.
METHOD: The process involves heating the long DNA strand and staple strands to 90°C-95°C, followed by gradual cooling to form the origami structure. Within this process, the degradation of the DNA triangle will be examined under the effects of pre-heating the M13 ssDNA to various temperatures, starting at 95°C, for different time intervals. Gel electrophoresis is used to confirm the formation of nanostructures while imaging with atomic force microscope (AFM) will show the morphology of the DNA triangles.
CONCLUSION: When a single-stranded DNA scaffold is exposed to high temperatures it may undergo heatinduced degradation, which results in improper formation of DNA origami triangles and reduced yield. This provides insight in determining the thermal stability of the long circular backbone for synthesizing DNA origami structures and improving overall yield.
Effects of Thermal Damage on the Formation of DNA Origami
INTRODUCTION: In DNA nanotechnology, using DNA as a building block in fabricating nanostructures has expanded the scope of DNA-focused research. With the DNA origami method, a long single-stranded DNA scaffold is folded into nanoscale shapes using complementary staple strands of oligonucleotides. As this technique is used for many biomedical applications such as drug delivery, the structural stability of the DNA origami is essential to designing more robust and reliable nanostructures for practical use. The aim of this study is to see the effects of heat damage on the construction of these DNA origami nanostructures, using DNA triangles as a model structure.
METHOD: The process involves heating the long DNA strand and staple strands to 90°C-95°C, followed by gradual cooling to form the origami structure. Within this process, the degradation of the DNA triangle will be examined under the effects of pre-heating the M13 ssDNA to various temperatures, starting at 95°C, for different time intervals. Gel electrophoresis is used to confirm the formation of nanostructures while imaging with atomic force microscope (AFM) will show the morphology of the DNA triangles.
CONCLUSION: When a single-stranded DNA scaffold is exposed to high temperatures it may undergo heatinduced degradation, which results in improper formation of DNA origami triangles and reduced yield. This provides insight in determining the thermal stability of the long circular backbone for synthesizing DNA origami structures and improving overall yield.