NAME¶
SiRNA - Perl object for designing small inhibitory RNAs.
SYNOPSIS¶
use Bio::Tools::SiRNA;
my $sirna_designer = Bio::Tools::SiRNA->new( -target => $bio_seq,
-rules => 'saigo'
);
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 ("\t", $pair->start, $pair->end, $pair->rank,
$sense_oligo_sequence, $antisense_oligo_sequence), "\n";
}
DESCRIPTION¶
Package for designing siRNA reagents.
Input is a Bio::SeqI-compliant object (the target).
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.
This verion 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.
In addition, this module provides three 'extra' rules which can be added above
and beyond any ruleset.
- 1.
- SiRNAs that overlap known SNPs (identified as SeqFeatures with primary tag
= variation) can be avoided.
- 2.
- 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.
- 3.
- SiRNAs may also be selected in the 3 prime UTR of a gene by setting
$sirna_designer-> include_3pr() to true.
EXPORT¶
None.
SEE ALSO¶
Bio::Tools::Run::Mdust, Bio::SeqFeature::SiRNA::Pair,
Bio::SeqFeature::SiRNA::Oligo..
FEEDBACK¶
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.
bioperl-l@bioperl.org - General discussion
http://bioperl.org/wiki/Mailing_lists - About the mailing lists
Support¶
Please direct usage questions or support issues to the mailing list:
bioperl-l@bioperl.org
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.
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:
https://github.com/bioperl/bioperl-live/issues
AUTHOR¶
Donald Jackson (donald.jackson@bms.com)
APPENDIX¶
The rest of the documentation details each of the object methods. Internal
methods are usually preceded with a _
new¶
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 'rank' 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().
target¶
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)
rules¶
Title : rules
Usage : $sirna->rules('ruleset')
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
design¶
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
add_oligos¶
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)