In Vitro Selection of Cancer-Specific RNA Aptamers

  • Lee Young-Ju (Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University) ;
  • Lee Seong-Wook (Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University)
  • 발행 : 2006.07.01

초록

In this study, nuclease-resistant RNA aptamers that are specific for Jurkat T leukemia cells were selected by a subtractive systemic evolution of ligands by exponential enrichment (SELEX) method. A randomized nuclease-resistant RNA library was incubated with normal peripheral blood mononuclear cells (PBMC) in each round to preclude RNAs that recognize the common cellular components on the surface of normal and cancer cells. The precluded RNAs were used for the selection of Jurkat T cell-specific aptamers, and the specific RNAs were then gradually enriched from start to the following selections. After 16 rounds of the subtractive SELEX, the selected aptamers were found to preferentially bind to Jurkat T cells, but not to the normal PBMC, evidenced by fluorescence-activated cell sorting analysis. Thus, the subtractive SELEX can be used to identify ligands to cancer-specific biological markers without prior knowledge of the nature of markers. The aptamers could be applied to specific cell sorting, tumor therapy, and diagnosis, and moreover, to find cancer cell-specific markers.

키워드

참고문헌

  1. Bae, S. J., J. H. Oum, S. Sharma, J. Park, and S. W. Lee. 2002. In vitro selection of specific RNA inhibitors of NFATc. Biochem. Biophys. Res. Commun. 298: 486-492 https://doi.org/10.1016/S0006-291X(02)02490-7
  2. Burgstaller, P., A. Girod, and M. Blind. 2002. Aptamers as tools for target prioritization and lead identification. Drug Discov. Today 7: 1221-1228 https://doi.org/10.1016/S1359-6446(02)02522-9
  3. Daniels, D. A., H. Chen, B. J. Ricke, K. M. Swiderek, and L. Gold. 2003. A tenascin-C aptamer identified by tumor cell SELEX: Systematic evolution of ligands by exponential enrichment. Proc. Natl. Acad. Sci. USA 100: 15416-15421
  4. Ellington, A. D. and J. W. Szostak. 1990. In vitro selection of RNA molecules that bind specific ligands. Nature 346: 818-822 https://doi.org/10.1038/346818a0
  5. Gold, L., P. Allen, J. Binkley, D. Brown, D. Schneider, S. R. Eddy, C. Tuerk, L. Green, S. Macdougal, and D. Tasset. 1993. RNA: The shape of things to come, pp. 497-510. In R. F. Gestelend and J. F. Atkins (eds.). The RNA World. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., U.S.A
  6. Hwang, B., K. Han, and S. W. Lee. 2003. Prevention of passively transferred experimental autoimmune myasthenia gravis by an in vitro selected RNA aptamer. FEBS Lett. 548: 85-89 https://doi.org/10.1016/S0014-5793(03)00745-2
  7. Hwang, B. and S. W. Lee. 2005. Analysis of in vivo interaction of HCV NS3 protein and specific RNA aptamer with yeast three-hybrid system. J. Microbiol. Biotechnol. 15: 660-664
  8. Jung, H. S. and S. W. Lee. 2005. Re-engineering of carcinoembryonic antigen RNA with the group I intron of Tetrahymena thermophila by targeted trans-splicing. J. Microbiol. Biotechnol. 15: 1408-1413
  9. Kim, H., S. J. Kim, S. N. Park, and J. W. Oh. 2004. Antiviral effect of amphotericin B on Japanese encephalitis virus replication. J. Microbiol. Biotechnol. 14: 121-127
  10. Klubmann, S., A. Nolte, A. Bald, V. A. Erdmann, and J. P. Furste. 1996. Mirror-image RNA that binds D-adenosine. Nat. Biotechnol.14: 1112-1115 https://doi.org/10.1038/nbt0996-1112
  11. Lauhon, C. T. and J. W. Szostak. 1995. RNA aptamers that bind flavin and nicotinamide redox cofactors. J. Am. Chem. Soc. 117: 1246-1257 https://doi.org/10.1021/ja00109a008
  12. Lee, S. W. and B. Sullenger. 1996. Isolation of a nuclease resistant decoy RNA that selectively blocks autoantibody binding to insulin receptors on human lymphocytes. J. Exp. Med.194: 315-324
  13. Lee, S. W. and B. Sullenger. 1997. Isolation of a nucleaseresistant decoy RNA that can protect human acetylcholine receptors from myasthenic antibodies. Nat. Biotechnol. 15: 41-45 https://doi.org/10.1038/nbt0197-41
  14. Lim, H., M. K. Kim, Y. H. Cho, J. M. Kim, Y. Lim, and C. H. Lee. 2004. Inhibition of cell cycle progression and induction of apoptosis in HeLa cells by HY558-1, novel CDK inhibitor isolated from Penicillium minioluteum F558. J. Microbiol. Biotechnol. 14: 978-984
  15. Lupold, S. E., B. Hicke, Y. Lin, and D. S. Coffey. 2002. Identification and characterization of nuclease-stabilized RNA molecules that bind human prostate cancer cells via the prostate-specific membrane antigen. Cancer Res. 62: 4029-4033
  16. Morris, K. N., K. B. Jensen, C. M. Julin, M. Well, and L. Gold. 1998. High affinity ligands from in vitro selection: Complex targets. Proc. Natl. Acad. Sci. USA 95: 2902-2907
  17. Pagratis, N. C., C. Bell, Y. F. Chang, S. Jennings, T. Fitzwater, D. Jellinek, and C. Dang. 1997. Potent 2'-amino, and 2'-fluoro-2'deoxyribonucleotide RNA inhibitors of keratinocyte growth factor. Nature Biotechnol. 15: 68-73 https://doi.org/10.1038/nbt0197-68
  18. Park, K. S., Y. J. Bae, E. J. Jung, and S. J. Kang. 2005. RTPCR-based detection of six garlic viruses and their phylogenetic relationships. J. Microbiol. Biotechnol. 15: 1110-1114
  19. Rosemeyer, V., A. Laubrock, and R. Seibl. 1995. Nonradioactive 3'-end-Iabeling of RNA molecules of different lengths by terminal deoxynucleotidyltransferase. Anal. Biochem. 224: 446-449 https://doi.org/10.1006/abio.1995.1068
  20. Rusconi, C. P., J. D. Roberts, G. A. Pitoc, S. M. Nimjee, R. R. White, G. Jr. Quick, E. Scardino, W. P. Fay, and B. A. Sullenger. 2004. Antidote-mediated control of an anticoagulant aptamer in vivo. Nat. Biotechnol. 22: 1423-1428 https://doi.org/10.1038/nbt1023
  21. Seo, H. S. and S. W. Lee. 2000. In vitro selection of the 2'-fluoro-2'-deoxyribonucleotide decoy RNA inhibitor of myasthenic autoantibodies. J. Microbiol. Biotechnol. 10: 707-713
  22. Strausberg, R. L., A. J. Simpson, L. J. Old, and G. J. Riggins. 2004. Oncogenomics and the development of new cancer therapies. Nature 429: 469-474 https://doi.org/10.1038/nature02627
  23. Sullenger, B. A. and E. Gilboa. 2002. Emerging clinical applications of RNA. Nature 418: 252-258 https://doi.org/10.1038/418252a
  24. Tuerk, C. and L. Gold. 1990. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249: 505-510 https://doi.org/10.1126/science.2200121
  25. Wang, C., M. Zhang, G. Yang, D. Zhang, H. Ding, H. Wang, M. Fan, B. Shen, and N. Shap. 2003. Single-stranded DNA aptamers that bind differentiated but not parental cells: Subtractive systemic evolution of ligands by exponential enrichment. J. Biotechnol. 102: 15-22 https://doi.org/10.1016/S0168-1656(02)00360-7