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Always turn off hyphenation; it makes .\" way too many mistakes in technical documents. .if n .ad l .nh .SH "NAME" SiRNA \- Perl object for designing small inhibitory RNAs. .SH "SYNOPSIS" .IX Header "SYNOPSIS" .Vb 1 \& use Bio::Tools::SiRNA; \& \& my $sirna_designer = Bio::Tools::SiRNA\->new( \-target => $bio_seq, \& \-rules => \*(Aqsaigo\*(Aq \& ); \& my @pairs = $sirna_designer\->design; \& \& foreach $pair (@pairs) { \& my $sense_oligo_sequence = $pair\->sense\->seq; \& my $antisense_oligo_sequence = $pair\->antisense\->seq; \& \& # print out results \& print join ("\et", $pair\->start, $pair\->end, $pair\->rank, \& $sense_oligo_sequence, $antisense_oligo_sequence), "\en"; \& } .Ve .SH "DESCRIPTION" .IX Header "DESCRIPTION" Package for designing siRNA reagents. .PP Input is a Bio::SeqI\-compliant object (the target). .PP Output is a list of Bio::SeqFeature::SiRNA::Pair objects, which are added to the feature table of the target sequence. Each Bio::SeqFeature::SiRNA::Pair contains two subfeatures (Bio::SeqFeature::Oligo objects) which correspond to the individual oligos. These objects provide accessors for the information on the individual reagent pairs. .PP This version of Bio::Tools::SiRNA represents a major change in architecture. Specific 'rulesets' for siRNA selection as developed by various groups are implemented as Bio::Tools::SiRNA::Ruleset objects, which inherit from Bio::Tools::SiRNA. This will make it easier to add new rule sets or modify existing approaches. Currently the Tuschl and Ui-Tei (2004) rules are implemented. For consistency, the Tuschl rules are implemented by default. .PP In addition, this module provides three 'extra' rules which can be added above and beyond any ruleset. .IP "1." 3 SiRNAs that overlap known SNPs (identified as SeqFeatures with primary tag = variation) can be avoided. .IP "2." 3 Other regions (with primary tag = 'Excluded') can also be skipped. I use this with Bio::Tools::Run::Mdust to avoid low-complexity regions (must be run separately), but other programs could also be used. .IP "3." 3 SiRNAs may also be selected in the 3 prime \s-1UTR\s0 of a gene by setting \&\f(CW$sirna_designer\fR\->\fIinclude_3pr()\fR to true. .SS "\s-1EXPORT\s0" .IX Subsection "EXPORT" None. .SH "SEE ALSO" .IX Header "SEE ALSO" Bio::Tools::Run::Mdust, Bio::SeqFeature::SiRNA::Pair, Bio::SeqFeature::SiRNA::Oligo.. .SH "FEEDBACK" .IX Header "FEEDBACK" .SS "Mailing Lists" .IX Subsection "Mailing Lists" User feedback is an integral part of the evolution of this and other Bioperl modules. Send your comments and suggestions preferably to the Bioperl mailing list. Your participation is much appreciated. .PP .Vb 2 \& bioperl\-l@bioperl.org \- General discussion \& http://bioperl.org/wiki/Mailing_lists \- About the mailing lists .Ve .SS "Support" .IX Subsection "Support" Please direct usage questions or support issues to the mailing list: .PP \&\fIbioperl\-l@bioperl.org\fR .PP rather than to the module maintainer directly. Many experienced and reponsive experts will be able look at the problem and quickly address it. Please include a thorough description of the problem with code and data examples if at all possible. .SS "Reporting Bugs" .IX Subsection "Reporting Bugs" Report bugs to the Bioperl bug tracking system to help us keep track of the bugs and their resolution. Bug reports can be submitted via the web: .PP .Vb 1 \& https://github.com/bioperl/bioperl\-live/issues .Ve .SH "AUTHOR" .IX Header "AUTHOR" Donald Jackson (donald.jackson@bms.com) .SH "APPENDIX" .IX Header "APPENDIX" The rest of the documentation details each of the object methods. Internal methods are usually preceded with a _ .SS "new" .IX Subsection "new" .Vb 10 \& Title : new \& Usage : my $sirna_designer = Bio::Tools::SiRNA\->new(); \& Function : Constructor for designer object \& Returns : Bio::Tools::SiRNA object \& Args : target \- the target sequence for the SiRNAs as a Bio::Seq::RichSeq \& start_pad \- distance from the CDS start to skip (default 75) \& end_pad \- distance from the CDS end to skip (default 50) \& include_3pr \- set to true to include SiRNAs in the 3prime UTR (default false) \& rules \- rules for selecting siRNAs, currently supporting saigo and tuschl \& min_gc \- minimum GC fraction (NOT percent) (default 0.4) \& max_gc \- maximum GC fraction (NOT percent) (default 0.6) \& cutoff \- worst \*(Aqrank\*(Aq accepted(default 3) \& avoid_snps \- boolean \- reject oligos that overlap a variation \& SeqFeature in the target (default true) \& gstring \- maximum allowed consecutive Gs. \& Too many can cause problems in synthesis (default 4) \& Note : All arguments can also be changed/accessed using autoloaded \& methods such as: \& \& my $start_pad = $sirna_designer\->start_pad(). .Ve .SS "target" .IX Subsection "target" .Vb 7 \& Title : target \& Usage : my $target_seq = $sirna_designer\->target(); # get the current target \& OR \& $sirna_designer\->target($new_target_seq); # set a new target \& Function : Set/get the target as a Bio::SeqI\-compliant object \& Returns : a Bio::SeqI\-compliant object \& Args : a Bio::SeqI\-compliant object (optional) .Ve .SS "rules" .IX Subsection "rules" .Vb 6 \& Title : rules \& Usage : $sirna\->rules(\*(Aqruleset\*(Aq) \& Purpose : set/get ruleset to use for selecting SiRNA oligo pairs. \& Returns : not sure yet \& Args : a ruleset name (currently supported: Tuschl, Saigo) \& or a Bio::Tools::SiRNA::RulesetI compliant object .Ve .SS "design" .IX Subsection "design" .Vb 5 \& Title : design \& Usage : my @pairs = $sirna_designer\->design(); \& Purpose : Design SiRNA oligo pairs. \& Returns : A list of SiRNA pairs as Bio::SeqFeature::SiRNA::Pair objects \& Args : none .Ve .SS "add_oligos" .IX Subsection "add_oligos" .Vb 4 \& Title : add_oligos \& Usage : $sirna_designer\->add_oligos($sequence, $start, $rank); \& Purpose : Add SiRNA olgos to target Bio::Seq as Bio::SeqFeature::SiRNA::Pair objects \& Args : Oligo sequence and start position (required), rank/score (optional) .Ve