Abstract
Small interfering RNAs (siRNAs) have been widely exploited for nucleotide-sequence-specific posttranscriptional gene silencing, as a tool to investigate gene function in eukaryotes, and they hold promise as potential therapeutic agents. Conventionally designed siRNAs are 21-mers with symmetric 2-nt 3′ overhangs that mimic intermediates (microRNAs or miRNAs) of the natural processing of longer dsRNA (double-stranded RNA). siRNAs are sequences with full c omplementarity to their target mRNA and can be generated by either chemical synthesis or processing of shRNAs (short hairpin RNAs) transcribed from DNA vectors. To minimize off-target effects, any homology to nontarget mRNA can be verified using the expressed sequence tag (EST) database for the relevant organism. Here, we provide a practical guide and an overview to the design and selection of effective and specifc siRNAs.
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Acknowledgments
This work was supported by Yorkshire Cancer Research (YCR) and Prostate Cancer Research Foundation (PCRF).
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Dawson, L.A., Usmani, B.A. (2008). Design, Manufacture, and Assay of the Efficacy of siRNAs for Gene Silencing. In: Starkey, M., Elaswarapu, R. (eds) Genomics Protocols. Methods in Molecular Biology™, vol 439. Humana Press. https://doi.org/10.1007/978-1-59745-188-8_27
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DOI: https://doi.org/10.1007/978-1-59745-188-8_27
Publisher Name: Humana Press
Print ISBN: 978-1-58829-871-3
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