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Delivering siRNAs to Difficult Cell Types

Electroporation of Primary, Neuronal and Other Hard-to-transfect Cells

Dmitriy Ovcharenko, Rich Jarvis, Kevin Kelnar, David Brown, Ambion, Inc.


RNA interference (RNAi) is a powerful experimental tool for reducing the expression of specific genes. It is used routinely for gene function analysis, target validation, and gene discovery. RNAi is also exploited to create new gene-specific therapeutics. Scientists performing RNAi experiments in mammalian systems must deliver siRNAs into cells where the siRNAs guide the RNA-induced silencing complex (RISC) to target mRNA for cleavage. Lipid-based transfection reagents are typically used for siRNA delivery in immortalized cell lines. However these reagents tend to be inefficient for siRNA delivery to most primary and neuronal cell types and to cells grown in suspension. Here we demonstrate the use of electroporation to deliver siRNAs into primary cells and other hard-to-transfect cell types. Ambion's siPORT siRNA Electroporation Buffer can be used with commonly available electroporators to ensure highly efficient siRNA delivery into cells while maintaining high levels of cell viability.


An Alternative to Lipid-based Transfection
Mammalian cells can be successfully loaded with exogenous siRNA when the correct method and matrix of transfection conditions are employed. Chemical transfection (e.g. using lipid-based reagents) is used routinely to del
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