More complete gene silencing by fewer siRNAs: Transparent optimized design and biophysical signature

Istvan Ladunga

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Highly accurate knockdown functional analyses based on RNA interference (RNAi) require the possible most complete hydrolysis of the targeted mRNA while avoiding the degradation of untargeted genes (off-target effects). This in turn requires significant improvements to target selection for two reasons. First, the average silencing activity of randomly selected siRNAs is as low as 62%. Second, applying more than five different siRNAs may lead to saturation of the RNA-induced silencing complex (RISC) and to the degradation of untargeted genes. Therefore, selecting a small number of highly active siRNAs is critical for maximizing knockdown and minimizing off-target effects. To satisfy these needs, a publicly available and transparent machine learning tool is presented that ranks all possible siRNAs for each targeted gene. Support vector machines (SVMs) with polynomial kernels and constrained optimization models select and utilize the most predictive effective combinations from 572 sequence, thermodynamic, accessibility and self-hairpin features over 2200 published siRNAs. This tool reaches an accuracy of 92.3% in cross-validation experiments. We fully present the underlying biophysical signature that involves free energy, accessibility and dinucleotide characteristics. We show that while complete silencing is possible at certain structured target sites, accessibility information improves the prediction of the 90% active siRNA target sites. Fast siRNA activity predictions can be performed on our web server at http://optirna.unl.edu/.

Original languageEnglish (US)
Pages (from-to)433-440
Number of pages8
JournalNucleic acids research
Volume35
Issue number2
DOIs
StatePublished - Jan 1 2007

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Gene Silencing
Small Interfering RNA
RNA-Induced Silencing Complex
Genes
RNA Interference
Thermodynamics
Hydrolysis
Messenger RNA

ASJC Scopus subject areas

  • Genetics

Cite this

More complete gene silencing by fewer siRNAs : Transparent optimized design and biophysical signature. / Ladunga, Istvan.

In: Nucleic acids research, Vol. 35, No. 2, 01.01.2007, p. 433-440.

Research output: Contribution to journalArticle

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