SLIDE 1
OVERVIEW
- 1. INTRODUCTION
NANYANG RESEARCH PROGRAMME -SPMS07 DNA Origami Assembled by DNA - - PowerPoint PPT Presentation
NANYANG RESEARCH PROGRAMME -SPMS07 DNA Origami Assembled by DNA - - PowerPoint PPT Presentation
NANYANG RESEARCH PROGRAMME -SPMS07 DNA Origami Assembled by DNA Dendrimers for Drug Delivery Liu Haoyun Temasek Junior College OVERVIEW 1. INTRODUCTION 2.AIM 3.METHEDOLOGY 4.DISCUSSION OF RESULTS 5.CONCLUSION 1. INTRODUCTION 1.1 DNA
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SLIDE 3
- 1. INTRODUCTION
1.1 DNA DENDRIMERS
Definition:
- Molecules formed by the attachment of DNA molecules
to a typically symmetrical core
- Basic building blocks of the DNA scaffold in DNA
hydrogels
SLIDE 4
- 1. INTRODUCTION
1.2 DNA HYDROGELS
Definition
- Polymeric networks formed by DNA dendrimers
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- 1. INTRODUCTION
1.2 DNA HYDROGELS
Characteristics:
- Bio-compatibility
- Molecular recognition capacity
- Nano-scale control capability
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- 1. INTRODUCTION
1.2 DNA HYDROGELS
Applications:
- Bio-sensing
- 3D growing medium
- Specific drug delivery
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- 2. AIM
To synthesize DNA dendrimers and assemble DNA nano-hydrogels that could potentially be used in specific drug delivery
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- 3. METHODOLOGY
3.1 SYNTHESIS OF DENDRITIC DNA 3.2 PURIFICATION OF DENDRITIC DNA 3.3 SELF-ASSEMBLY OF DNA NANOHYDROGEL
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- 3. METHODOLOGY
3.1 SYNTHESIS OF DENDRITIC DNA
Synthesis of DNA using long trebler phosphoamidite as the branching reagent through Bioautomation Mermaid 4 via solid phase oligonucleotide synthesis Synthesized dendritic DNA consisted of 1 loading branch for functionalizing and 3 gelation branches for hybridization
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- 3. METHODOLOGY
3.1 SYNTHESIS OF DENDRITIC DNA
Cleavage of DNA molecules from the controlled pore glass via incubation in 1ml of 33% NH4OH solution at 25°C for 10 minutes Deprotection of the DNA through controlled heating at 60°C in a dry bath
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- 3. METHODOLOGY
3.2 PURIFICATION OF DENDRITIC DNA
Purification of the dendritic DNA was achieved through Reverse Phase HPLC with polar phase being 0.1 M TEAA pH7 and non-polar phase ACN DNA was collected in TEAA
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- 3. METHODOLOGY
3.2 PURIFICATION OF DENDRITIC DNA
Dendritic DNA was incubated in 200µl of 80% acetic acid for 15 minutes for the cleavage of DMT group Second round of HPLC to separate pure dendritic DNA from the DMT group.
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- 3. METHODOLOGY
3.3 SELF-ASSEMBLY OF DNA NANOHYDROGEL
200nM purified dendritic DNA was mixed with 200nM
- f linker DNA
The mixture was heated in 95°C dry bath for 10 minutes for DNA annealing and cooled down naturally for self-assembly Resultant DNA nanohydrogels collected and concentrated though centrifugation
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- 4. DISCUSSION OF RESULTS
4.1 DNA DENDRIMER SYNTHESIS
Sequence of synthesized dendritic DNA : (CGA TTA CAG CTT GCT)3 D TTTCGA TCG Molecular weight derived from MALDI-TOF showed a reading of 16640.73
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- 4. DISCUSSION OF RESULTS
4.1 DNA DENDRIMER SYNTHESIS
Theoretical molecular weight was 16945.16 Synthesized DNA dendrimers were usable
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- 4. DISCUSSION OF RESULTS
4.2 DNA NANO HYDROGEL SYNTHESIS
Synthesized DNA nanohydrogels was examined under AFM to determine their size An average size was108.853nm
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- 4. DISCUSSION OF RESULTS
4.2 DNA NANO HYDROGEL SYNTHESIS
Data collected from AFM verified through DLS The readings collected from DLS showed an average size of 129.1nm possibly due to the fact that theDNA nanohydrogel was in vacuum-driedstate in AFM
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- 5. CONCLUSION
The DNA nanohydrogels were synthesized successfully from DNA dendrimers and are ready for loading of Doxorubicin for drug test
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