Title Page
Abstract
Contents
Chapter 1. Circular Antisense Oligonucleotide for RNA degradation 8
1.1. Abstract 9
1.2. Introduction 9
1.3. Materials and Methods 11
1.4. Results and Discussion 15
1.5. Conclusion 20
Chapter 2. U-rich nanoparticle for cell proliferation inhibition 29
2.1. Abstract 30
2.2. Introduction 30
2.3. Materials and Method 31
2.4. Results and Discussion 35
2.5. Conclusion 40
References 54
Abstract (Korean) 58
Figure 1.1. (A) Schematic illustration of circular ASO synthesis. (B) Gel electrophoresis to investigate the cyclization of linear ASO through DNA ligase.... 22
Figure 1.2. (A) Gel electrophoresis of linear ASO and circular ASO with or without Exonuclease I. (B) Enzymatic stability test of linear ASO and circular... 23
Figure 1.3. (A) Cell viability and (B) Relative luciferase activity of HeLa Firefly luciferase treated with various ASOs. Relative luciferase activity was... 24
Figure 1.4. Schematic illustration of (A) RNA granule synthesis via Rolling Circle Transcription (RCT) and (B) RNA granule degradation model with C-ASO. 25
Figure 1.5. Gel electrophoresis of RNA granule degradation depending on circular ASO concentration. (0, 5, 25, 100, 300, 500 nM) 26
Figure 1.6. (A) AFM image and height profile of untreated (UT), negative control (N.C), and circular ASO (C-ASO) groups. Scale: 20 μm. (Negative... 27
Figure 1.7. (A) AFM image and height profile of untreated (UT), negative control (N.C), and circular ASO (C-ASO) groups. Scale: 2 μm. (Negative... 28
Figure 2.1. (A) SEM image, (B) EDX-mapping images, and (C) TEM image of UNP. (D) Size and zeta potential of UNP measured using Dynamic Light... 42
Figure 2.2. Confocal microscope and Z-stack confocal laser scanning to confirm HeLa cell intracellular uptake of (A) Cy5-labelled -LF UNP and (B)... 43
Figure 2.3. (A) Cytometry analysis of HeLa cells with untreated (UT), Cy5-labelled -LF UNP, and Cy5-labelled +LF UNP after incubation with 24 h. (B)... 44
Figure 2.4. HeLa cells were treated with (A) UNP or (B) R-NP for 4, 12, 24, 48 h. CCK-8 assay was used to measure the viability of cervical cancer cells.... 45
Figure 2.5. Fluorescence microscopy images of Live/Dead assay of HeLa cells treated with R-NP and UNP for 48 h at 10 μg/mL concentration. Green signals... 46
Figure 2.6. (A) Cell proliferation assay image and (B) relative cell proliferation rate of HeLa GFP (left) co-cultured with HDF(right): Scale bar=100 µm. The... 47
Figure 2.7. (A) Schematic illustration of mRNA extraction from HeLa cells and (B) relative total mRNA amount of HeLa cell after mRNA separation. The... 49
Figure 2.8. qRT-PCR Cycle Threshold (Cq) values depicting the expression levels of key genes-GAPDH, Bcl-2, and cMYC-in HeLa cells. Data are... 50
Figure 2.9. Tumor suppression of UNP in HeLa xenograft model. (A) In vivo fluorescence images are shown injected with PBS, 25 μg of UNP, 100 μg UNP... 51
Figure 2.10. Tumor suppression of UNP in HeLa xenograft model. (A) In vivo fluorescence images are shown injected with UNP after 0.5, 1, 2, 6 h... 53