sIR: siRNA Information Resource, a web-based tool for siRNA sequence design and analysis and an open access siRNA database

BackgroundRNA interference has revolutionized our ability to study the effects of altering the expression of single genes in mammalian (and other) cells through targeted knockdown of gene expression. In this report we describe a web-based computational tool, siRNA Information Resource (sIR), which consists of a new open source database that contains validation information about published siRNA sequences and also provides a user-friendly interface to design and analyze siRNA sequences against a chosen target sequence.ResultsThe siRNA design tool described in this paper employs empirically determined rules derived from a meta-analysis of the published data; it uses a weighted scoring system that determines the optimal sequence within a target mRNA and thus aids in the rational selection of siRNA sequences. This scoring system shows a non-linear correlation with the knockdown efficiency of siRNAs. sIR provides a fast, customized BLAST output for all selected siRNA sequences against a variety of databases so that the user can verify the uniqueness of the design. We have pre-designed siRNAs for all the known human genes (24,502) in the Refseq database. These siRNAs were pre-BLASTed against the human Unigene database to estimate the target specificity and all results are available online.ConclusionAlthough most of the rules for this scoring system were influenced by previously published rules, the weighted scoring system provides better flexibility in designing an appropriate siRNA when compared to the un-weighted scoring system. sIR is not only a comprehensive tool used to design siRNA sequences and lookup pre-designed siRNAs, but it is also a platform where researchers can share information on siRNA design and use.

[1]  Michael Zuker,et al.  Mfold web server for nucleic acid folding and hybridization prediction , 2003, Nucleic Acids Res..

[2]  M. Amarzguioui,et al.  Positional effects of short interfering RNAs targeting the human coagulation trigger Tissue Factor. , 2002, Nucleic acids research.

[3]  David Botstein,et al.  SOURCE: a unified genomic resource of functional annotations, ontologies, and gene expression data , 2003, Nucleic Acids Res..

[4]  Laura Mariani,et al.  The energy profiling of short interfering RNAs is highly predictive of their activity. , 2004, Oligonucleotides.

[5]  A. Reynolds,et al.  Rational siRNA design for RNA interference , 2004, Nature Biotechnology.

[6]  T. Tuschl,et al.  Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate , 2001, The EMBO journal.

[7]  K. Ui-Tei,et al.  Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference. , 2004, Nucleic acids research.

[8]  G. Sczakiel,et al.  Local RNA target structure influences siRNA efficacy: a systematic global analysis. , 2005, Journal of molecular biology.

[9]  T. Tuschl,et al.  RNA Interference and Small Interfering RNAs , 2001, Chembiochem : a European journal of chemical biology.

[10]  Xin Zheng,et al.  BIOINFORMATICSAPPLICATIONS NOTE doi:10.1093/bioinformatics/btl026 Databases and ontologies , 2005 .

[11]  N. Schütze siRNA technology , 2004, Molecular and Cellular Endocrinology.

[12]  Szymon M. Kielbasa,et al.  HuSiDa—the human siRNA database: an open-access database for published functional siRNA sequences and technical details of efficient transfer into recipient cells , 2004, Nucleic Acids Res..

[13]  D. Dykxhoorn,et al.  Killing the messenger: short RNAs that silence gene expression , 2003, Nature Reviews Molecular Cell Biology.

[14]  Dieter Huesken,et al.  Design of a genome-wide siRNA library using an artificial neural network , 2005, Nature Biotechnology.

[15]  Anastasia Khvorova,et al.  3′ UTR seed matches, but not overall identity, are associated with RNAi off-targets , 2006, Nature Methods.

[16]  Alan M Gewirtz,et al.  Progress in the Development of Nucleic Acids Therapeutics for Cancer , 2004, Cancer biology & therapy.

[17]  M. Amarzguioui,et al.  An algorithm for selection of functional siRNA sequences. , 2004, Biochemical and biophysical research communications.

[18]  A. Fire,et al.  Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans , 1998, Nature.

[19]  Thomas Tuschl,et al.  siRNAs: applications in functional genomics and potential as therapeutics , 2004, Nature Reviews Drug Discovery.