請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/47397
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 白果能(Konan Peck) | |
dc.contributor.author | Te - Kai Ting | en |
dc.contributor.author | 丁德楷 | zh_TW |
dc.date.accessioned | 2021-06-15T05:58:01Z | - |
dc.date.available | 2013-10-07 | |
dc.date.copyright | 2011-10-07 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-18 | |
dc.identifier.citation | Alexopoulou, L., Holt, A. C., Medzhitov, R., and Flavell, R. A. (2001). Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3. Nature 413, 732-738.
Amarzguioui, M., Lundberg, P., Cantin, E., Hagstrom, J., Behlke, M. A., and Rossi, J. J. (2006). Rational design and in vitro and in vivo delivery of Dicer substrate siRNA. Nat Protoc 1, 508-517. Amarzguioui, M., and Prydz, H. (2004). An algorithm for selection of functional siRNA sequences. Biochem Biophys Res Commun 316, 1050-1058. Amarzguioui, M., and Rossi, J. J. (2008). Principles of Dicer substrate (D-siRNA) design and function. Methods Mol Biol 442, 3-10. Angaji, S. A., Hedayati, S. S., Poor, R. H., Madani, S., Poor, S. S., and Panahi, S. (2010). Application of RNA interference in treating human diseases. J Genet 89, 527-537. Asagiri, M., Hirai, T., Kunigami, T., Kamano, S., Gober, H. J., Okamoto, K., Nishikawa, K., Latz, E., Golenbock, D. T., Aoki, K., et al. (2008). Cathepsin K-dependent toll-like receptor 9 signaling revealed in experimental arthritis. Science 319, 624-627. Barrat, F. J., Meeker, T., Gregorio, J., Chan, J. H., Uematsu, S., Akira, S., Chang, B., Duramad, O., and Coffman, R. L. (2005). Nucleic acids of mammalian origin can act as endogenous ligands for Toll-like receptors and may promote systemic lupus erythematosus. J Exp Med 202, 1131-1139. Basu, S., Pathak, S. K., Chatterjee, G., Pathak, S., Basu, J., and Kundu, M. (2008). Helicobacter pylori protein HP0175 transactivates epidermal growth factor receptor through TLR4 in gastric epithelial cells. J Biol Chem 283, 32369-32376. Berkhout, B., and Haasnoot, J. (2006). The interplay between virus infection and the cellular RNA interference machinery. FEBS Lett 580, 2896-2902. Blander, J. M., and Medzhitov, R. (2004). Regulation of phagosome maturation by signals from toll-like receptors. Science 304, 1014-1018. Boone, D. L., and Ma, A. (2003). Connecting the dots from Toll-like receptors to innate immune cells and inflammatory bowel disease. J Clin Invest 111, 1284-1286. Bourquin, C., Anz, D., Zwiorek, K., Lanz, A. L., Fuchs, S., Weigel, S., Wurzenberger, C., von der Borch, P., Golic, M., Moder, S., et al. (2008). Targeting CpG oligonucleotides to the lymph node by nanoparticles elicits efficient antitumoral immunity. J Immunol 181, 2990-2998. Boutros, M., Kiger, A. A., Armknecht, S., Kerr, K., Hild, M., Koch, B., Haas, S. A., Paro, R., and Perrimon, N. (2004). Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science 303, 832-835. Brightbill, H. D., Libraty, D. H., Krutzik, S. R., Yang, R. B., Belisle, J. T., Bleharski, J. R., Maitland, M., Norgard, M. V., Plevy, S. E., Smale, S. T., et al. (1999). Host defense mechanisms triggered by microbial lipoproteins through toll-like receptors. Science 285, 732-736. Bulut, Y., Faure, E., Thomas, L., Equils, O., and Arditi, M. (2001). Cooperation of Toll-like receptor 2 and 6 for cellular activation by soluble tuberculosis factor and Borrelia burgdorferi outer surface protein A lipoprotein: role of Toll-interacting protein and IL-1 receptor signaling molecules in Toll-like receptor 2 signaling. J Immunol 167, 987-994. Bumcrot, D., Manoharan, M., Koteliansky, V., and Sah, D. W. (2006). RNAi therapeutics: a potential new class of pharmaceutical drugs. Nat Chem Biol 2, 711-719. Burdelya, L. G., Krivokrysenko, V. I., Tallant, T. C., Strom, E., Gleiberman, A. S., Gupta, D., Kurnasov, O. V., Fort, F. L., Osterman, A. L., Didonato, J. A., et al. (2008). An agonist of toll-like receptor 5 has radioprotective activity in mouse and primate models. Science 320, 226-230. Cekaite, L., Furset, G., Hovig, E., and Sioud, M. (2007). Gene expression analysis in blood cells in response to unmodified and 2'-modified siRNAs reveals TLR-dependent and independent effects. J Mol Biol 365, 90-108. Chang, Y. C., Kao, W. C., Wang, W. Y., Yang, R. B., and Peck, K. (2009). Identification and characterization of oligonucleotides that inhibit Toll-like receptor 2-associated immune responses. FASEB J 23, 3078-3088. Chuang, T., and Ulevitch, R. J. (2001). Identification of hTLR10: a novel human Toll-like receptor preferentially expressed in immune cells. Biochim Biophys Acta 1518, 157-161. Clevers, H. (2004). At the crossroads of inflammation and cancer. Cell 118, 671-674. Cox, J. C., and Ellington, A. D. (2001). Automated selection of anti-protein aptamers. Bioorg Med Chem 9, 2525-2531. Cristofaro, P., and Opal, S. M. (2003). The Toll-like receptors and their role in septic shock. Expert Opin Ther Targets 7, 603-612. Cullen, B. R. (2006). Is RNA interference involved in intrinsic antiviral immunity in mammals? Nat Immunol 7, 563-567. Dassie, J. P., Liu, X. Y., Thomas, G. S., Whitaker, R. M., Thiel, K. W., Stockdale, K. R., Meyerholz, D. K., McCaffrey, A. P., McNamara, J. O., 2nd, and Giangrande, P. H. (2009). Systemic administration of optimized aptamer-siRNA chimeras promotes regression of PSMA-expressing tumors. Nat Biotechnol 27, 839-849. Dua, P., Kim, S., and Lee, D. K. (2008). Patents on SELEX and therapeutic aptamers. Recent Pat DNA Gene Seq 2, 172-186. Ellington, A. D., and Szostak, J. W. (1990). In vitro selection of RNA molecules that bind specific ligands. Nature 346, 818-822. Fedorov, Y., Anderson, E. M., Birmingham, A., Reynolds, A., Karpilow, J., Robinson, K., Leake, D., Marshall, W. S., and Khvorova, A. (2006). Off-target effects by siRNA can induce toxic phenotype. RNA 12, 1188-1196. Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., and Mello, C. C. (1998). Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806-811. Fukata, M., Chen, A., Vamadevan, A. S., Cohen, J., Breglio, K., Krishnareddy, S., Hsu, D., Xu, R., Harpaz, N., Dannenberg, A. J., et al. (2007). Toll-like receptor-4 promotes the development of colitis-associated colorectal tumors. Gastroenterology 133, 1869-1881. Hamdy, S., Molavi, O., Ma, Z., Haddadi, A., Alshamsan, A., Gobti, Z., Elhasi, S., Samuel, J., and Lavasanifar, A. (2008). Co-delivery of cancer-associated antigen and Toll-like receptor 4 ligand in PLGA nanoparticles induces potent CD8+ T cell-mediated anti-tumor immunity. Vaccine 26, 5046-5057. Hayashi, E. A., Akira, S., and Nobrega, A. (2005). Role of TLR in B cell development: signaling through TLR4 promotes B cell maturation and is inhibited by TLR2. J Immunol 174, 6639-6647. Hayashi, F., Smith, K. D., Ozinsky, A., Hawn, T. R., Yi, E. C., Goodlett, D. R., Eng, J. K., Akira, S., Underhill, D. M., and Aderem, A. (2001). The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410, 1099-1103. He, W., Liu, Q., Wang, L., Chen, W., Li, N., and Cao, X. (2007). TLR4 signaling promotes immune escape of human lung cancer cells by inducing immunosuppressive cytokines and apoptosis resistance. Mol Immunol 44, 2850-2859. Heil, F., Hemmi, H., Hochrein, H., Ampenberger, F., Kirschning, C., Akira, S., Lipford, G., Wagner, H., and Bauer, S. (2004). Species-specific recognition of single-stranded RNA via toll-like receptor 7 and 8. Science 303, 1526-1529. Hemmi, H., Takeuchi, O., Kawai, T., Kaisho, T., Sato, S., Sanjo, H., Matsumoto, M., Hoshino, K., Wagner, H., Takeda, K., and Akira, S. (2000). A Toll-like receptor recognizes bacterial DNA. Nature 408, 740-745. Hertz, C. J., Kiertscher, S. M., Godowski, P. J., Bouis, D. A., Norgard, M. V., Roth, M. D., and Modlin, R. L. (2001). Microbial lipopeptides stimulate dendritic cell maturation via Toll-like receptor 2. J Immunol 166, 2444-2450. Heymann, W. R. (2006). Toll-like receptors in acne vulgaris. J Am Acad Dermatol 55, 691-692. Hybarger, G., Bynum, J., Williams, R. F., Valdes, J. J., and Chambers, J. P. (2006). A microfluidic SELEX prototype. Anal Bioanal Chem 384, 191-198. Iwasaki, A., and Medzhitov, R. (2004). Toll-like receptor control of the adaptive immune responses. Nat Immunol 5, 987-995. Jeon, S. H., Kayhan, B., Ben-Yedidia, T., and Arnon, R. (2004). A DNA aptamer prevents influenza infection by blocking the receptor binding region of the viral hemagglutinin. J Biol Chem 279, 48410-48419. Johnson, G. B., Brunn, G. J., and Platt, J. L. (2003). Activation of mammalian Toll-like receptors by endogenous agonists. Crit Rev Immunol 23, 15-44. Kamath, R. S., and Ahringer, J. (2003). Genome-wide RNAi screening in Caenorhabditis elegans. Methods 30, 313-321. Kim, H. S., Han, M. S., Chung, K. W., Kim, S., Kim, E., Kim, M. J., Jang, E., Lee, H. A., Youn, J., Akira, S., and Lee, M. S. (2007). Toll-like receptor 2 senses beta-cell death and contributes to the initiation of autoimmune diabetes. Immunity 27, 321-333. Knuefermann, P., Schwederski, M., Velten, M., Krings, P., Ehrentraut, H., Rudiger, M., Boehm, O., Fink, K., Dreiner, U., Grohe, C., et al. (2008). Bacterial DNA induces myocardial inflammation and reduces cardiomyocyte contractility: role of toll-like receptor 9. Cardiovasc Res 78, 26-35. Lemaitre, B., Nicolas, E., Michaut, L., Reichhart, J. M., and Hoffmann, J. A. (1996). The dorsoventral regulatory gene cassette spatzle/Toll/cactus controls the potent antifungal response in Drosophila adults. Cell 86, 973-983. Levy-Nissenbaum, E., Radovic-Moreno, A. F., Wang, A. Z., Langer, R., and Farokhzad, O. C. (2008). Nanotechnology and aptamers: applications in drug delivery. Trends Biotechnol 26, 442-449. Liu, X., Ukai, T., Yumoto, H., Davey, M., Goswami, S., Gibson, F. C., 3rd, and Genco, C. A. (2008). Toll-like receptor 2 plays a critical role in the progression of atherosclerosis that is independent of dietary lipids. Atherosclerosis 196, 146-154. Long, S. B., Long, M. B., White, R. R., and Sullenger, B. A. (2008). Crystal structure of an RNA aptamer bound to thrombin. RNA 14, 2504-2512. Lu, R., Maduro, M., Li, F., Li, H. W., Broitman-Maduro, G., Li, W. X., and Ding, S. W. (2005). Animal virus replication and RNAi-mediated antiviral silencing in Caenorhabditis elegans. Nature 436, 1040-1043. Matsuda, N., and Hattori, Y. (2006). Systemic inflammatory response syndrome (SIRS): molecular pathophysiology and gene therapy. J Pharmacol Sci 101, 189-198. McNamara, J. O., 2nd, Andrechek, E. R., Wang, Y., Viles, K. D., Rempel, R. E., Gilboa, E., Sullenger, B. A., and Giangrande, P. H. (2006). Cell type-specific delivery of siRNAs with aptamer-siRNA chimeras. Nat Biotechnol 24, 1005-1015. Medzhitov, R. (2001). Toll-like receptors and innate immunity. Nat Rev Immunol 1, 135-145. Medzhitov, R., Preston-Hurlburt, P., and Janeway, C. A., Jr. (1997). A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature 388, 394-397. Meng, G., Rutz, M., Schiemann, M., Metzger, J., Grabiec, A., Schwandner, R., Luppa, P. B., Ebel, F., Busch, D. H., Bauer, S., et al. (2004). Antagonistic antibody prevents toll-like receptor 2-driven lethal shock-like syndromes. J Clin Invest 113, 1473-1481. Mi, J., Liu, Y., Rabbani, Z. N., Yang, Z., Urban, J. H., Sullenger, B. A., and Clary, B. M. (2010). In vivo selection of tumor-targeting RNA motifs. Nat Chem Biol 6, 22-24. Mosing, R. K., Mendonsa, S. D., and Bowser, M. T. (2005). Capillary electrophoresis-SELEX selection of aptamers with affinity for HIV-1 reverse transcriptase. Anal Chem 77, 6107-6112. Nakamura, M., Shimizu, Y., Sato, Y., Miyazaki, Y., Satoh, T., Mizuno, M., Kato, Y., Hosaka, Y., and Furusako, S. (2007). Toll-like receptor 4 signal transduction inhibitor, M62812, suppresses endothelial cell and leukocyte activation and prevents lethal septic shock in mice. Eur J Pharmacol 569, 237-243. Napoli, C., Lemieux, C., and Jorgensen, R. (1990). Introduction of a Chimeric Chalcone Synthase Gene into Petunia Results in Reversible Co-Suppression of Homologous Genes in trans. Plant Cell 2, 279-289. Nomura, N., Miyajima, N., Sazuka, T., Tanaka, A., Kawarabayasi, Y., Sato, S., Nagase, T., Seki, N., Ishikawa, K., and Tabata, S. (1994). Prediction of the coding sequences of unidentified human genes. I. The coding sequences of 40 new genes (KIAA0001-KIAA0040) deduced by analysis of randomly sampled cDNA clones from human immature myeloid cell line KG-1 (supplement). DNA Res 1, 47-56. Obonyo, M., Sabet, M., Cole, S. P., Ebmeyer, J., Uematsu, S., Akira, S., and Guiney, D. G. (2007). Deficiencies of myeloid differentiation factor 88, Toll-like receptor 2 (TLR2), or TLR4 produce specific defects in macrophage cytokine secretion induced by Helicobacter pylori. Infect Immun 75, 2408-2414. Okamoto, M., and Sato, M. (2003). Toll-like receptor signaling in anti-cancer immunity. J Med Invest 50, 9-24. Padmanabhan, K., and Tulinsky, A. (1996). An ambiguous structure of a DNA 15-mer thrombin complex. Acta Crystallogr D Biol Crystallogr 52, 272-282. Palauqui, J. C., Elmayan, T., Pollien, J. M., and Vaucheret, H. (1997). Systemic acquired silencing: transgene-specific post-transcriptional silencing is transmitted by grafting from silenced stocks to non-silenced scions. EMBO J 16, 4738-4745. Pecot, C. V., Calin, G. A., Coleman, R. L., Lopez-Berestein, G., and Sood, A. K. (2011). RNA interference in the clinic: challenges and future directions. Nat Rev Cancer 11, 59-67. Pei, Z., Lin, D., Song, X., Li, H., and Yao, H. (2008). TLR4 signaling promotes the expression of VEGF and TGFbeta1 in human prostate epithelial PC3 cells induced by lipopolysaccharide. Cell Immunol 254, 20-27. Phillips, J. A., Lopez-Colon, D., Zhu, Z., Xu, Y., and Tan, W. (2008). Applications of aptamers in cancer cell biology. Anal Chim Acta 621, 101-108. Prinz, M., Garbe, F., Schmidt, H., Mildner, A., Gutcher, I., Wolter, K., Piesche, M., Schroers, R., Weiss, E., Kirschning, C. J., et al. (2006). Innate immunity mediated by TLR9 modulates pathogenicity in an animal model of multiple sclerosis. J Clin Invest 116, 456-464. Qiu, S., Adema, C. M., and Lane, T. (2005). A computational study of off-target effects of RNA interference. Nucleic Acids Res 33, 1834-1847. Ramirez Cruz, N. E., Maldonado Bernal, C., Cuevas Uriostegui, M. L., Castanon, J., Lopez Macias, C., and Isibasi, A. (2004). Toll-like receptors: dysregulation in vivo in patients with acute respiratory distress syndrome. Rev Alerg Mex 51, 210-217. Rifkin, I. R., Leadbetter, E. A., Busconi, L., Viglianti, G., and Marshak-Rothstein, A. (2005). Toll-like receptors, endogenous ligands, and systemic autoimmune disease. Immunol Rev 204, 27-42. Rodriguez, D., Keller, A. C., Faquim-Mauro, E. L., de Macedo, M. S., Cunha, F. Q., Lefort, J., Vargaftig, B. B., and Russo, M. (2003). Bacterial lipopolysaccharide signaling through Toll-like receptor 4 suppresses asthma-like responses via nitric oxide synthase 2 activity. J Immunol 171, 1001-1008. Romano, N., and Macino, G. (1992). Quelling: transient inactivation of gene expression in Neurospora crassa by transformation with homologous sequences. Mol Microbiol 6, 3343-3353. Sanjuan, M. A., Dillon, C. P., Tait, S. W., Moshiach, S., Dorsey, F., Connell, S., Komatsu, M., Tanaka, K., Cleveland, J. L., Withoff, S., and Green, D. R. (2007). Toll-like receptor signalling in macrophages links the autophagy pathway to phagocytosis. Nature 450, 1253-1257. Sarir, H., Henricks, P. A., van Houwelingen, A. H., Nijkamp, F. P., and Folkerts, G. (2008). Cells, mediators and Toll-like receptors in COPD. Eur J Pharmacol 585, 346-353. Schutz, S., and Sarnow, P. (2006). Interaction of viruses with the mammalian RNA interference pathway. Virology 344, 151-157. Seya, T., Akazawa, T., Uehori, J., Matsumoto, M., Azuma, I., and Toyoshima, K. (2003). Role of toll-like receptors and their adaptors in adjuvant immunotherapy for cancer. Anticancer Res 23, 4369-4376. Shangguan, D., Cao, Z. C., Li, Y., and Tan, W. (2007). Aptamers evolved from cultured cancer cells reveal molecular differences of cancer cells in patient samples. Clin Chem 53, 1153-1155. Shim, M. S., and Kwon, Y. J. (2010). Efficient and targeted delivery of siRNA in vivo. FEBS J 277, 4814-4827. Sioud, M. (2011). Promises and challenges in developing RNAi as a research tool and therapy. Methods Mol Biol 703, 173-187. Stram, Y., and Kuzntzova, L. (2006). Inhibition of viruses by RNA interference. Virus Genes 32, 299-306. Sun, J., Walsh, M., Villarino, A. V., Cervi, L., Hunter, C. A., Choi, Y., and Pearce, E. J. (2005). TLR ligands can activate dendritic cells to provide a MyD88-dependent negative signal for Th2 cell development. J Immunol 174, 742-751. Syed, M. A., and Pervaiz, S. (2010). Advances in aptamers. Oligonucleotides 20, 215-224. Tasset, D. M., Kubik, M. F., and Steiner, W. (1997). Oligonucleotide inhibitors of human thrombin that bind distinct epitopes. J Mol Biol 272, 688-698. Tuerk, C., and Gold, L. (1990). Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249, 505-510. Vogl, T., Tenbrock, K., Ludwig, S., Leukert, N., Ehrhardt, C., van Zoelen, M. A., Nacken, W., Foell, D., van der Poll, T., Sorg, C., and Roth, J. (2007). Mrp8 and Mrp14 are endogenous activators of Toll-like receptor 4, promoting lethal, endotoxin-induced shock. Nat Med 13, 1042-1049. Wang, X. H., Aliyari, R., Li, W. X., Li, H. W., Kim, K., Carthew, R., Atkinson, P., and Ding, S. W. (2006). RNA interference directs innate immunity against viruses in adult Drosophila. Science 312, 452-454. Whitehead, K. A., Langer, R., and Anderson, D. G. (2009). Knocking down barriers: advances in siRNA delivery. Nat Rev Drug Discov 8, 129-138. Wilkins, C., Dishongh, R., Moore, S. C., Whitt, M. A., Chow, M., and Machaca, K. (2005). RNA interference is an antiviral defence mechanism in Caenorhabditis elegans. Nature 436, 1044-1047. Wyllie, D. H., Kiss-Toth, E., Visintin, A., Smith, S. C., Boussouf, S., Segal, D. M., Duff, G. W., and Dower, S. K. (2000). Evidence for an accessory protein function for Toll-like receptor 1 in anti-bacterial responses. J Immunol 165, 7125-7132. Xu, Y., Tao, X., Shen, B., Horng, T., Medzhitov, R., Manley, J. L., and Tong, L. (2000). Structural basis for signal transduction by the Toll/interleukin-1 receptor domains. Nature 408, 111-115. Zambon, R. A., Vakharia, V. N., and Wu, L. P. (2006). RNAi is an antiviral immune response against a dsRNA virus in Drosophila melanogaster. Cell Microbiol 8, 880-889. Zhou, J., Li, H., Li, S., Zaia, J., and Rossi, J. J. (2008). Novel dual inhibitory function aptamer-siRNA delivery system for HIV-1 therapy. Mol Ther 16, 1481-1489. Zhou, J., and Rossi, J. J. (2011a). Aptamer-targeted RNAi for HIV-1 therapy. Methods Mol Biol 721, 355-371. Zhou, J., and Rossi, J. J. (2011b). Cell-specific aptamer-mediated targeted drug delivery. Oligonucleotides 21, 1-10. Zhou, J., Shu, Y., Guo, P., Smith, D. D., and Rossi, J. J. (2011). Dual functional RNA nanoparticles containing phi29 motor pRNA and anti-gp120 aptamer for cell-type specific delivery and HIV-1 Inhibition. Methods. Zhou, J., Swiderski, P., Li, H., Zhang, J., Neff, C. P., Akkina, R., and Rossi, J. J. (2009). Selection, characterization and application of new RNA HIV gp 120 aptamers for facile delivery of Dicer substrate siRNAs into HIV infected cells. Nucleic Acids Res 37, 3094-3109. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/47397 | - |
dc.description.abstract | TLR 接受器在先天免疫反應中占有重要的一環。主要會辨識外來病原如細菌與病毒。這些病原具有稱為病原相關分子結構(Pathogen associated molecular pattern, PAMP)的特殊結構。 在一般狀況下,這些病原相關分子結構會刺激TLR接受器使其活化進而使免疫細胞分泌大量的細胞激素(Cytokines),產生發炎反應,以消滅入侵的病原菌。 雖然TLR 接受器在人體對抗外來疾病佔有非常重要的角色,但近期研究中發現,在TLR接受器過度活化下,將造成許多臨床症狀的產生,例如關節炎(arthritis)、動脈硬化(arteriosclerosis)、癌症甚至是敗血症的發生。因此發展能夠調節TLR 接受器功能的藥物將是一個十分重要的目標。適體是由一段DNA 、RNA分子所構成的特殊序列結構,因為DNA 、RNA分子會因序列的不同而產生各式各樣可能的二級或三級結構,這些特殊結構經由SELEX (Systematic evolution of ligands by exponential enrichment)方式篩選,便可篩選出會與目標物有專一性結合的序列,由於舊式SELEX方式對於篩選針對細胞膜上的目標物具有一定的困難,因此本論文利用三種SELEX方法,分別是IP-SELEX、cell-based SELEX以及endocytic SELEX來篩選對TLR2接受器具專一性並可藉此進入細胞的適體。實驗結果成功獲得專一辨識並拮抗TLR2接受器之適體。將適體與siRNA連結後,適體可以將siRNA帶進細胞內,並更進一步抑制LTA所造成的NF-κB活性。本篇研究提供一個完整的實驗流程方法,從如何篩選針對細胞膜上TLR2接受器並進入細胞中之適體,進一步找出具有功能的適體序列並對適體做定量以及定性的分析,最後將適體接上siRNA進行免疫反應抑制實驗。同樣的流程亦可套用在其他TLR接受器上,配合接上抑制不同發炎相關基因的siRNA,來達到調節免疫反應的作用,這些適體與siRNA的嵌合體在針對發炎相關疾病的治療上,具有相當大的潛力。 | zh_TW |
dc.description.abstract | Toll-like receptors (TLRs) play important roles in the innate immunity against
invading microorganisms. They recognize pathogen-associated molecular patterns (PAMPs) that are expressed on pathogens, and mediate the production of cytokines necessary for triggering effective immunity. Although TLRs are critical for immune responses, it can cause severe clinical manifestation such as arthritis, atherosclerosis, cancer, or even sepsis while it’s over activated. Identification of TLRs antagonists is therefore considered as a promising direction in the regulation of TLRs associated inflammatory disorder. Aptamers are single-stranded RNA or DNA molecules isolated through an in vitro selection process. In this study, I use a selection strategy that combines immunopreciptate (IP) SELEX, cell-based SELEX and endocytic SELEX. This modification facilitates the screening of high-affinity aptamers against human TLR2, which can further be engulfed by TLR2-expressing cells. After 16 rounds of selection, the aptamers against TLR2 with the ability to trigger endocytosis were identified. My data suggested that the isolated aptamers were effectively engulfed through TLR2-dependent pathway. These aptamers were further analyzed and characterized to find out if they could antagonize TLR2 receptor by using NF-κB reporter assays. Several isolated aptamers exhibited strong inhibition of LTA-induced TLR2 response. Moreover, I had conjugated the isolated aptamer with p65 siRNA to further knockdown the NF-κB signal induced by TLR2 independent pathway. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T05:58:01Z (GMT). No. of bitstreams: 1 ntu-100-R98445117-1.pdf: 2872639 bytes, checksum: adb329b55e63a0f46e738e6d0f61ae2c (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 誌謝 i
目錄 iii 圖目錄 v 表目錄 vii 中文摘要 viii ABSTRACT ix 第一章 前言 1 第二章 文獻回顧 3 2.1 適體簡介 3 2.2 TLRs簡介 5 2.3 TLR在免疫系統中扮演的角色 6 2.4 siRNA簡介 7 2.5 RNAi的機制 8 2.6 siRNA的應用 9 2.7 適體-siRNA嵌合體(chimera) 10 第三章 材料與方法 11 3.1細胞株與試劑 11 3.2引子與適體 11 3.3 IP-SELEX 12 3.4 Cell-based SELEX 13 3.5 Endocytic SELEX 13 3.6 適體定序 14 3.7 檢測試體是否具有抑制TLR2接受器之活性 14 3.8 共軛焦螢光顯微鏡成像 14 3.9 流式細胞儀分析適體結合之專一性 15 3.10 適體胞吞測試 15 3.11 適體與siRNA的結合 15 第四章 實驗結果 17 4.1 辨識TLR2的DNA 適體 17 4.2適體對TLR2的專一性 18 4.3 endocytosis的能力 18 4.4檢測試體是否具有抑制TLR2接受器之活性 19 4.5適體與siRNA嵌合體 20 第五章 討論 21 5.1 本篇篩選方式之優點 21 5.2適體結合訊號與胞吞作用的相關性 21 5.3所篩選到的適體對TLR2的抑制效果 22 5.4同時選用人類與老鼠細胞來篩選TLR2適體的原因 22 5.5適體-siRNA 嵌合物 23 5.6適體-siRNA 嵌合物是否具有副作用 24 5.7抑制實驗結果不如預期的原因探討 24 5.8 未來可繼續發展的方向 25 第六章 結論 26 第七章 表與圖 27 第八章 參考文獻: 46 第九章 附錄 57 | |
dc.language.iso | zh-TW | |
dc.title | TLR2適體-干擾RNA嵌合物於抑制免疫之研究 | zh_TW |
dc.title | Identification and characterization of TLR2 aptamer for siRNA delivery to suppress immune responses | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 楊瑞彬(Ruey-Bing Yang),楊淑美(Shu-Mei Liang) | |
dc.subject.keyword | 適體,TLR接受器,SELEX,發炎反應,適體與siRNA複合體, | zh_TW |
dc.subject.keyword | aptamer,TLR2,SELEX,inflammation,aptamer-siRNA chimera, | en |
dc.relation.page | 70 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-19 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-100-1.pdf 目前未授權公開取用 | 2.81 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。