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  • Title: Biodegradable polymer-mediated sh/siRNA delivery for cancer studies.
    Author: Jere DJ, Cho CS.
    Journal: Methods Mol Biol; 2010; 623():243-69. PubMed ID: 20217556.
    Abstract:
    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 (beta-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.
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