Biodegradable Polymer-Mediated sh/siRNA Delivery for Cancer Studies
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 wordssiRNA shRNA Nonviral vector Polymeric carrier Gene delivery siRNA delivery Lung cancer siRNA protocols Cancer protocols
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