Biodegradable Polymer-Mediated sh/siRNA Delivery for Cancer Studies

  • Dhananjay J. Jere
  • Chong-Su ChoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 623)


Discovery of RNA interference (RNAi)-mediated specific gene silencing has raised hope for cancer therapy. Unfortunately, the execution of RNAi by delivering small-interfering RNA (siRNA) or small hairpin RNA (shRNA) remains a prime challenge. A methodical evaluation of cationic polymers in RNAi-based cancer studies may offer a promising solution to this problem. In this chapter, we report the methodologies for comprehensive characterization of a biodegradable polymeric system for sh/siRNA delivery in cancer studies. The chapter will describe synthesis, characterization, and optimization of biodegradable poly (β-amino ester) for sh/siRNA delivery. The protocols are provided for shRNA and siRNA complex preparation, stability and morphology study. Also, detailed methods are provided for the intracellular tracking and transfection of sh/siRNA using polymeric carrier. In addition, step-wise information is provided for the in vitro silencing of oncoprotein to study important cancer properties, including proliferation, malignancy, and metastasis of cancer cells.

Key words

siRNA shRNA Nonviral vector Polymeric carrier Gene delivery siRNA delivery Lung cancer siRNA protocols Cancer protocols 


  1. 1.
    Amarzguioui, M., Rossi, J.J. and Kim, D. (2005) Approaches for chemically synthesized siR NA and vector-mediated RNAi. FEBS Lett. 579, 5974-5981.CrossRefPubMedGoogle Scholar
  2. 2.
    Dykxhoorn, D.M. and Lieberman, J. (2006) Knocking down disease with siRNAs. Cell 126, 231-235.CrossRefPubMedGoogle Scholar
  3. 3.
    Engelke, D.R. and Rossi, J.J. (eds.) (2005) RNA interference. Elsevier, San Diego, California, USA.Google Scholar
  4. 4.
    Devi, ­­­G.R. (2006) siRNA-based approaches in cancer therapy. Cancer Gene Ther. 13, 819-829.CrossRefPubMedGoogle Scholar
  5. 5.
    Pai, S.I., Lin, Y.Y., Macaes, B., Meneshian, A., Hung, C.F. and Wu, T.C. (2006) Prospects of RNA interference therapy for cancer. Gene Ther. 13, 464-477.CrossRefPubMedGoogle Scholar
  6. 6.
    Jere, D., Xu, C.X., Arote, R., Yun, C.H., Cho, M.H. and Cho, C.S. (2008) Poly(beta-amino ester) as a carrier for si/shRNA delivery in lung cancer cells. Biomaterials 29, 2535-2547.CrossRefPubMedGoogle Scholar
  7. 7.
    Xu, C.X., Jere, D., Jin, H., Chang, S.H., Chung, Y.S., Shin, J.Y., et al. (2008) Poly(ester amine)-mediated, aerosol-delivered Akt1 small interfering RNA suppresses lung tumorigenesis. Am. J. Respir. Crit. Care Med., 178, 60-73.CrossRefPubMedGoogle Scholar
  8. 8.
    Kim, T.H., Jiang, H.L., Jere, D., Park, I.K. and Cho, C.S. (2007) Chemical modification of chitosan as a gene carrier in vitro and in vivo. Prog. Polym. Sci. 32, 726-753.CrossRefGoogle Scholar
  9. 9.
    Gary, D.J., Puri, N. and Won, Y.Y. (2007) Polymer-based siRNA delivery: perspectives on the fundamental and phenomenological distinctions from polymer-based DNA delivery. J. Control Release, 121, 64-73.CrossRefPubMedGoogle Scholar
  10. 10.
    Jere, D., Yoo, M.K., Arote, R., Kim, T.H., Cho, M.H., Nah, J.W., Choi, Y.J. and Cho, C.S. (2007) Poly (amino ester) composed of poly (ethylene glycol) and aminosilane prepared by combinatorial chemistry as a gene carrier. Pharm. Res., 25, 875-885.CrossRefPubMedGoogle Scholar
  11. 11.
    Jere, D., Kim, T.H., Arote, R., Jiang, H.L., Cho, M.H., Nah, J.W. and Cho, C.S. (2007) A Poly(β-amino ester) of spermine and poly(ethylene glycol) diacrylate as a gene carrier. Key Eng. Mater. 342-343, 425-428.CrossRefGoogle Scholar
  12. 12.
    Arote, R.B., Hwang, S.K., Yoo, M.K., Jere, D., Jiang, H.L., Kim, Y.K., et al. (2008) Biodegradable poly(ester amine) based on glycerol dimethacrylate and polyethylenimine as a gene carrier. J. Gene Med., 10, 1223-1235.CrossRefPubMedGoogle Scholar
  13. 13.
    Arote, R., Kim, T.H., Kim, Y.K., Hwang, S.K., Jiang, H.L., Song, H.H., et al. (2007) A biodegradable poly(ester amine) based on polycaprolactone and polyethylenimine as a gene carrier. Biomaterials 28, 735-744.CrossRefPubMedGoogle Scholar
  14. 14.
    Jere, D., Arote, R., Cho, M.H. and Cho, C.S. (2008) Biodegradable poly(B-amino ester) derivatives for gene and siRNA delivery. NOVA, New York.Google Scholar
  15. 15.
    Mather, B.D., Vishwanathan, K., Miller, K.M. and Long T.E., (2006) Michael addition reactions in macromolecular design for emerging technologies. Prog. Polym. Sci. 31, 487-531.CrossRefGoogle Scholar
  16. 16.
    Akinc, A., Anderson, D.G., Lynn, D.M. and Langer, R. (2003) Synthesis of poly(beta-amino ester)s optimized for highly effective gene delivery. Bioconjug. Chem. 14, 979-988.CrossRefPubMedGoogle Scholar
  17. 17.
    Anderson, D.G., Lynn, D.M. and Langer, R. (2003) Semi-automated synthesis and screening of a large library of degradable cationic polymers for gene delivery. Angew. Chem. Int. Ed. Engl. 42, 3153-3158.CrossRefPubMedGoogle Scholar
  18. 18.
    Park, M.R., Han, K.O., Han, I.K., Cho, M.H., Nah, J.W., Choi, Y.J. and Cho, C.S. (2005) Degradable polyethylenimine-alt-poly(ethylene glycol) copolymers as novel gene carriers. J. Control Release 105, 367-380.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of Agricultural BiotechnologyCollege of Agriculture and Life Sciences, Seoul National UniversitySeoulSouth Korea

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