請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/30753
標題: Effect of Tet-controlled RNAi system on the gene expression in Drosophila Schneider 2 (S2) cell line
四環黴素調控RNAi系統對果蠅S2細胞基因表現之影響
作者: 林憶淳
Lin, Yi-Chun
關鍵字: 四環黴素調控系統
http://etds.lib.nchu.edu.tw/etdservice/view_metadata?etdun=U0005-0802201109331200
RNAi
S2細胞
出版社: 昆蟲學系所
引用: Aagaard, L., M. Amarzguioui, G. Sun, L. C. Santos, A. Ehsani, H. Prydz, and J. J. Rossi. 2007. A facile lentiviral vector system for expression of doxycycline-inducible shRNAs: knockdown of the pre-miRNA processing enzyme Drosha. Mol Ther. 15: 938-945. Alphey, L. 2002. Re-engineering the sterile insect technique. Insect Biochem Mol Biol. 32: 1243-1247. Alphey, L., and M. Andreasen. 2002. Dominant lethality and insect population control. Mol Biochem Parasitol. 121: 173-178. Alphey, L., D. Nimmo, S. O''Connell, and N. Alphey. 2008. Insect population suppression using engineered insects. Adv Exp Med Biol. 627: 93-103. Benedict, M. Q., and A. S. Robinson. 2003. The first releases of transgenic mosquitoes: an argument for the sterile insect technique. Trends Parasitol. 19: 349-355. Berens, C., and W. Hillen. 2003. Gene regulation by tetracyclines. Constraints of resistance regulation in bacteria shape TetR for application in eukaryotes. Eur J Biochem. 270: 3109-3121. Bergmann, A., J. Agapite, K. McCall, and H. Steller. 1998. The Drosophila gene hid is a direct molecular target of Ras-dependent survival signaling. Cell. 95: 331-341. Cheikh, M., J. F. Howell, E. J. Harris, H. B. Salah, and F. Soria. 1975. Suppression of the mediterranean fruit fly in Tunisia with released sterile insects. J Econ Entomol. 68: 237-243. Chen, S. L., C. Cheng, and K. H. Lu. 2010. Cloning, characterization and promoter activity of the yolk protein gene, Bdyp1, in the oriental fruit fly Bactrocera dorsalis. Formosan Entomol. 30: 9-25. Collins, S. R., C. W. Weldon, C. Banos, and P. W. Taylor. 2009. Optimizing irradiation dose for sterility induction and quality of Bactrocera tryoni. J Econ Entomol. 102: 1791-1800. Curtis, C. F., G. D. Brooks, M. A. Ansari, K. K. Grover, B. S. Krishnamurthy, P. K. Rajagopalan, L. S. Sharma, V. P. Sharma, D. Singh, K. R. P. Singh, and M. Yasuno. 1982. A field trial on control of Culex quinquefasciatus by release of males of a strain integrating cytoplasmic incompatibility and a translocation. Ent exp & appl. 31: 181-190. Du, Q., H. Thonberg, J. Wang, C. Wahlestedt, and Z. Liang. 2005. A systematic analysis of the silencing effects of an active siRNA at all single-nucleotide mismatched target sites. Nucleic Acids Res. 33: 1671-1677. Elbashir, S. M., J. Harborth, W. Lendeckel, A. Yalcin, K. Weber, and T. Tuschl. 2001. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 411: 494-498. Fire, A., S. Xu, M. K. Montgomery, S. A. Kostas, S. E. Driver, and C. C. Mello. 1998. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 391: 806-811. Forstemann, K., M. D. Horwich, L. Wee, Y. Tomari, and P. D. Zamore. 2007. Drosophila microRNAs are sorted into functionally distinct argonaute complexes after production by dicer-1. Cell. 130: 287-297. Fu, G., K. C. Condon, M. J. Epton, P. Gong, L. Jin, G. C. Condon, N. I. Morrison, T. H. Dafa''alla, and L. Alphey. 2007. Female-specific insect lethality engineered using alternative splicing. Nat Biotechnol. 25: 353-357. Fu, G., R. S. Lees, D. Nimmo, D. Aw, L. Jin, P. Gray, T. U. Berendonk, H. White-Cooper, S. Scaife, H. Kim Phuc, O. Marinotti, N. Jasinskiene, A. A. James, and L. Alphey. 2010. Female-specific flightless phenotype for mosquito control. Proc Natl Acad Sci U S A. 107: 4550-4554. Gong, P., M. J. Epton, G. Fu, S. Scaife, A. Hiscox, K. C. Condon, G. C. Condon, N. I. Morrison, D. W. Kelly, T. Dafa''alla, P. G. Coleman, and L. Alphey. 2005. A dominant lethal genetic system for autocidal control of the Mediterranean fruitfly. Nat Biotechnol. 23: 453-456. Gossen, M., and H. Bujard. 1992. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. 89: 5547-5551. Hannon, G. J. 2002. RNA interference. Nature. 418: 244-251. Heinrich, J. C., and M. J. Scott. 2000. A repressible female-specific lethal genetic system for making transgenic insect strains suitable for a sterile-release program. Proc Natl Acad Sci U S A. 97: 8229-8232. Jeanson-Leh, L., J. Blondeau, and A. Galy. 2007. Optimization of short hairpin RNA for lentiviral-mediated RNAi against WAS. Biochem Biophys Res Commun. 362: 498-503. Kim, H. J., C. Gatz, W. Hillen, and T. R. Jones. 1995. Tetracycline repressor-regulated gene repression in recombinant human cytomegalovirus. J Virol. 69: 2565-2573. Knipling, E. F. 1959. Sterile-male method of population control. Science. 130: 902-904. Kutter, C., and P. Svoboda. 2008. miRNA, siRNA, piRNA: Knowns of the unknown. RNA Biol. 5: 181-188. Lang, Z., and J. M. Feingold. 1996. An autonomously replicating eukaryotic expression vector with a tetracycline-responsive promoter. Gene. 168: 169-171. Leung, R. K., and P. A. Whittaker. 2005. RNA interference: from gene silencing to gene-specific therapeutics. Pharmacol Ther. 107: 222-239. Lin, H., J. McGrath, P. Wang, and T. Lee. 2007. Cellular toxicity induced by SRF-mediated transcriptional squelching. Toxicol Sci. 96: 83-91. Markaki, M., D. Drabek, I. Livadaras, R. K. Craig, F. Grosveld, and C. Savakis. 2007. Stable expression of human growth hormone over 50 generations in transgenic insect larvae. Transgenic Res. 16: 99-107. Martinez, J., A. Patkaniowska, H. Urlaub, R. Luhrmann, and T. Tuschl. 2002. Single-stranded antisense siRNAs guide target RNA cleavage in RNAi. Cell. 110: 563-574. McDonald, I. C. 1971. A male-producing strain of the house fly. Science. 172: 489. McInnis, D., L. Leblanc, and R. Mau. 2007. Melon Fly (Diptera: Tephritidae) Genetic Sexing: All-male Sterile Fly Releases in Hawaii. Proc Hawaii Entomol Soc. 39: 105-110. Papathanos, P. A., H. C. Bossin, M. Q. Benedict, F. Catteruccia, C. A. Malcolm, L. Alphey, and A. Crisanti. 2009. Sex separation strategies: past experience and new approaches. Malar J. 8 Suppl 2: S5. Peters, L., and G. Meister. 2007. Argonaute proteins: mediators of RNA silencing. Mol Cell. 26: 611-623. Phuc, H. K., M. H. Andreasen, R. S. Burton, C. Vass, M. J. Epton, G. Pape, G. Fu, K. C. Condon, S. Scaife, C. A. Donnelly, P. G. Coleman, H. White-Cooper, and L. Alphey. 2007. Late-acting dominant lethal genetic systems and mosquito control. BMC Biol. 5: 11. Preall, J. B., Z. He, J. M. Gorra, and E. J. Sontheimer. 2006. Short interfering RNA strand selection is independent of dsRNA processing polarity during RNAi in Drosophila. Curr Biol. 16: 530-535. Rössler, Y. 1979. Auto mated sexing of Ceratitis capitata (Dip. :Tephritidae): The development of strains with inherited, sex-limited pupal color dimophism. Entomophaga. 24: 411. Reynolds, A., D. Leake, Q. Boese, S. Scaringe, W. S. Marshall, and A. Khvorova. 2004. Rational siRNA design for RNA interference. Nat Biotechnol. 22: 326-330. Ronaldson, E., and M. Bownes. 1995. Two independent cis-acting elements regulate the sex- and tissue-specific expression of yp3 in Drosophila melanogaster. Genet Res. 66: 9-17. Sadowski, I., J. Ma, S. Triezenberg, and M. Ptashne. 1988. GAL4-VP16 is an unusually potent transcriptional activator. Nature. 335: 563-564. Saul, S. H. 1990. A genetic sexing system to improve the sterile insect technique against the Mediterranean fruit fly. J Hered. 81: 75-78. Schetelig, M. F., C. Caceres, A. Zacharopoulou, G. Franz, and E. A. Wimmer. 2009. Conditional embryonic lethality to improve the sterile insect technique in Ceratitis capitata (Diptera: Tephritidae). BMC Biol. 7: 4. Schwarz, D. S., G. Hutvagner, T. Du, Z. Xu, N. Aronin, and P. D. Zamore. 2003. Asymmetry in the assembly of the RNAi enzyme complex. Cell. 115: 199-208. Seawright, J. A., P. E. Kaiser, D. A. Dame, and C. S. Lofgren. 1978. Genetic method for the preferential elimination of females of anopheles albimanus. Science. 200: 1303-1304. Shah, C., and K. Forstemann. 2008. Monitoring miRNA-mediated silencing in Drosophila melanogaster S2-cells. Biochim Biophys Acta. 1779: 766-772. Steiner, L. F., E. J. Harris, M. S. Fujimoto, and L. D. Christenson. 1965. Melon fly eradication by overflooding with sterile flies. J Econ Entomol. 58: 519-522. Strathdee, C. A., M. R. McLeod, and J. R. Hall. 1999. Efficient control of tetracycline-responsive gene expression from an autoregulated bi-directional expression vector. Gene. 229: 21-29. Thomas, D. D., C. A. Donnelly, R. J. Wood, and L. S. Alphey. 2000. Insect population control using a dominant, repressible, lethal genetic system. Science. 287: 2474-2476. Triezenberg, S. J., R. C. Kingsbury, and S. L. McKnight. 1988a. Functional dissection of VP16, the trans-activator of herpes simplex virus immediate early gene expression. Genes Dev. 2: 718-729. Triezenberg, S. J., K. L. LaMarco, and S. L. McKnight. 1988b. Evidence of DNA: protein interactions that mediate HSV-1 immediate early gene activation by VP16. Genes Dev. 2: 730-742. Vreysen, M. J., K. M. Saleh, M. Y. Ali, A. M. Abdulla, Z. R. Zhu, K. G. Juma, V. A. Dyck, A. R. Msangi, P. A. Mkonyi, and H. U. Feldmann. 2000. Glossina austeni (Diptera: Glossinidae) eradicated on the island of Unguja, Zanzibar, using the sterile insect technique. J Econ Entomol. 93: 123-135. Wakiyama, M., T. Matsumoto, and S. Yokoyama. 2005. Drosophila U6 promoter-driven short hairpin RNAs effectively induce RNA interference in Schneider 2 cells. Biochem Biophys Res Commun. 331: 1163-1170. Watson, J. D. 2007. Recombinant DNA: Gene and Genomes-a short course. pp. 219-246. In: R.M. Myers, A.A. Caudy, and J.A. Witkowski, eds. RNA interference regulates gene action. Cold spring harbor laboratory Press, New York. Wu, T. Y., L. Liono, S. L. Chen, C. Y. Chen, and Y. C. Chao. 2000. Expression of highly controllable genes in insect cells using a modified tetracycline-regulated gene expression system. J Biotechnol. 80: 75-83. Yang, D., H. Lu, and J. W. Erickson. 2000. Evidence that processed small dsRNAs may mediate sequence-specific mRNA degradation during RNAi in Drosophila embryos. Curr Biol. 10: 1191-1200.
摘要: 利用四環黴素調控系統(tetracycline-responsive expression system, TRES)表現以東方果實蠅(Bactrocera dorsalis (Hendel))肌動蛋白(actin)基因所設計的短髮夾核醣核酸(small hairpin RNA, shRNA)片段於果蠅Drosophlia Schneider 2 (S2) 細胞株,並測試TRES調控shRNA之表現及其對細胞之影響。本試驗構築之四環黴素調控系統,係以驅動(driver)載體上之東方果實蠅卵黃蛋白(yolk protein, yp)基因啟動子驅動四環黴素轉錄活化子(tetracycline-controllable transactivator, tTA)表現,再經由此tTA與表現(responder)載體上四環黴素操作子(tet operator)鍵結而促進後方shRNA片段表現;然而,tTA之活性可因加入四環黴素而被抑制,使之無法與其操作子結合而終止shRNA表現。卵黃蛋白啟動子僅於雌成蟲發育卵時受激素調控而活化,在S2細胞測試結果顯示此啟動子能被濃度為10-7-10-9 M 的20-羥基蛻皮激素(20-hydroxyecdysone, 20E)激活而表現tta 基因。然而,濃度高於10-7 M之20E雖可刺激tta大量表現,但細胞存活數卻明顯降低;此外,經雙轉染(co-transfection)驅動及表現載體之細胞,以10-8 M之20E處理後,可以偵測到shRNA片段的表現,顯示tTA與操作子結合後確實能轉錄出shRNA,顯示此系統在S2細胞中可以發揮作用,期望不久的未來能將此系統有效的應用於東方果實蠅蟲體內。
In this study, a tetracycline-responsive expression system (TRES) for expressing a small hairpin RNA (shRNA), i.e. part of actin gene of Bactrocera dorsalis (Hendel), was modified in Drosophila S2 cell line. This could be applied as an alternative method to control insect pests. For the driver construct of TRES, the promoter of yolk protein gene (yp) of B. dorsalis, was selected to drive the expression of tetracycline-controllable transactivator (tTA); and the expressed tTA proteins then bond to the tet operator on the responder construct of TRES to initiate the transcription of the DNA coding shRNA sequence; furthermore, the tTA activity could be inhibited by adding tetracycline, and under this circumstances, the shRNA expression was stopped. The yp promoter was only activated by hormone when the eggs started to develop in female adults, and proved to be activated under the stimulation of 10-7-10-9 M 20-hydroxyecdysone (20E) in the driver-transfected S2 cells. In the driver-transfected S2 cells, tta was significantly expressed by treating with 10-7 M, resulting in a large amount of cell death. In the cells co-transfected with both driver and responder, the shRNA expression was clearly detected after treating with 10-8 M 20E, suggesting that the tTA protein is able to activate the shRNA expression in S2 cells. In conclusion, it is demonstrated that this TRES is functional in S2 cells, and that, in the near future this system can further transferred into B. dorsalis to observe whether the system can function as well in this fly.
URI: http://hdl.handle.net/11455/30753
其他識別: U0005-0802201109331200
顯示於類別:昆蟲學系

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