Springer Protocols: Abstract: Design, Manufacture, and Assay of the Efficacy of siRNAs for Gene Silencing

Design, Manufacture, and Assay of the Efficacy of siRNAs for Gene Silencing

By: Louise A. Dawson⁴, Badar A. Usmani⁴

Abstract

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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.

Affiliation(s): (4) Institute of Cellular and Molecular Biology, University of Leeds, Leeds, UK

Book Title: Genomics Protocols

Series: Methods in Molecular Biology | Volume: 439 | Pub. Date: Feb-01-2008 | Page Range: 403-419 | DOI: 10.1007/978-1-59745-188-8_27

Subject: Genetics/Genomics

Key Words: small interfering RNA - gene silencing - endothelin-converting enzyme - prostate cancer - invasion - stromal-epithelial interactions

References

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