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Five Ways to Produce siRNAs

gene silencing can be readily cloned into a plasmid or viral vector to create an siRNA expression vector. The siRNA expression vector can then be used for stable expression and long term studies. One disadvantage of SECs is that they are not as easily transfected into cells as siRNAs. As new transfection agents and protocols are developed, however, SEC transfection efficiency should increase. Because they are generated by PCR, SECs are not amenable to scale up without being cloned into plasmids. Figure 3 shows data from an experiment in which SECs each containing one of three different promoters were tested for their ability to drive siRNA expression and induce knockdown of c-fos expression.

Best for:
Screening siRNA sequences and testing promoters before preparing vectors

Not suitable for:
Long term studies until cloned into vectors containing selectable markers

Ambion's Solution: Silencer Express siRNA Expression Cassettes
The Silencer Express Kits make it easy to produce siRNA expression cassettes (SECs) with a human H1, human U6, or mouse U6 promoter by PCR for use in RNAi studies. Because a single siRNA template oligonucleotide can be used with any of the three Silencer Express Kits, different promoters can be tested for their capacity to express active siRNAs in a specific cell line. Ambion's Silencer Express Kits provide the rea
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TRIS-EDTA based buffer prepared from RNase-free water suitable for storage and dilution of siRNA, pH 8, store at 4 C ...
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