# # = bio/util/sirna.rb - Class for designing small inhibitory RNAs # # Copyright:: Copyright (C) 2004, 2005 # Itoshi NIKAIDO # License:: LGPL # # $Id: sirna.rb,v 1.7 2005/12/27 17:27:38 k Exp $ # # == Bio::SiRNA - Designing siRNA. # # This class implements the selection rules described by Kumiko Ui-Tei # et al. (2004) and Reynolds et al. (2004). # # == Example # # seq = Bio::Sequence::NA.new(ARGF.read) # # sirna = Bio::SiRNA.new(seq) # pairs = sirna.design # # pairs.each do |pair| # puts pair.report # shrna = Bio::SiRNA::ShRNA.new(pair) # shrna.design # puts shrna.report # # puts shrna.top_strand.dna # puts shrna.bottom_strand.dna # end # # == References # # * Kumiko Ui-Tei et al. Guidelines for the selection of highly effective # siRNA sequences for mammalian and chick RNA interference. # Nucl. Acids. Res. 2004 32: 936-948. # # * Angela Reynolds et al. Rational siRNA design for RNA interference. # Nature Biotech. 2004 22: 326-330. # #-- # # This library is free software; you can redistribute it and/or # modify it under the terms of the GNU Lesser General Public # License as published by the Free Software Foundation; either # version 2 of the License, or (at your option) any later version. # # This library is distributed in the hope that it will be useful, # but WITHOUT ANY WARRANTY; without even the implied warranty of # MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU # Lesser General Public License for more details. # # You should have received a copy of the GNU Lesser General Public # License along with this library; if not, write to the Free Software # Foundation, Inc., 59 Temple Place, Suite 330, Boston, MA 02111-1307 USA # #++ # require 'bio/sequence' module Bio # = Bio::SiRNA # Designing siRNA. # # This class implements the selection rules described by Kumiko Ui-Tei # et al. (2004) and Reynolds et al. (2004). class SiRNA # A parameter of size of antisense. attr_accessor :antisense_size # A parameter of maximal %GC. attr_accessor :max_gc_percent # A parameter of minimum %GC. attr_accessor :min_gc_percent # Input is a Bio::Sequence::NA object (the target sequence). # Output is a list of Bio::SiRNA::Pair object. def initialize(seq, antisense_size = 21, max_gc_percent = 60.0, min_gc_percent = 40.0) @seq = seq.rna! @pairs = Array.new @antisense_size = antisense_size @max_gc_percent = max_gc_percent @min_gc_percent = min_gc_percent end # Ui-Tei's rule. def uitei?(target) return false unless /^.{2}[GC]/i =~ target return false unless /[AU].{2}$/i =~ target return false if /[GC]{9}/i =~ target one_third = target.size * 1 / 3 start_pos = @target_size - one_third - 1 remain_seq = target.subseq(start_pos, @target_size - 2) gc_number = remain_seq.scan(/[AU]/i).size return false if gc_number < 5 return true end # Reynolds' rule. def reynolds?(target) return false if /[GC]{9}/i =~ target return false unless /^.{4}A.{6}U.{2}[AUC].{5}[AU].{2}$/i =~ target return true end # same as design('uitei'). def uitei design('uitei') end # same as design('reynolds'). def reynolds design('reynolds') end # rule can be one of 'uitei' (default) and 'reynolds'. def design(rule = 'uitei') @target_size = @antisense_size + 2 target_start = 0 @seq.window_search(@target_size) do |target| antisense = target.subseq(1, @target_size - 2).complement.rna sense = target.subseq(3, @target_size) target_start += 1 target_stop = target_start + @target_size antisense_gc_percent = antisense.gc_percent next if antisense_gc_percent > @max_gc_percent next if antisense_gc_percent < @min_gc_percent case rule when 'uitei' next unless uitei?(target) when 'reynolds' next unless reynolds?(target) else raise NotImplementedError end pair = Bio::SiRNA::Pair.new(target, sense, antisense, target_start, target_stop, rule, antisense_gc_percent) @pairs.push(pair) end return @pairs end # = Bio::SiRNA::Pair class Pair attr_accessor :target attr_accessor :sense attr_accessor :antisense attr_accessor :start attr_accessor :stop attr_accessor :rule attr_accessor :gc_percent def initialize(target, sense, antisense, start, stop, rule, gc_percent) @target = target @sense = sense @antisense = antisense @start = start @stop = stop @rule = rule @gc_percent = gc_percent end # human readable report def report report = "### siRNA\n" report << 'Start: ' + @start.to_s + "\n" report << 'Stop: ' + @stop.to_s + "\n" report << 'Rule: ' + @rule.to_s + "\n" report << 'GC %: ' + @gc_percent.to_s + "\n" report << 'Target: ' + @target.upcase + "\n" report << 'Sense: ' + ' ' + @sense.upcase + "\n" report << 'Antisense: ' + @antisense.reverse.upcase + "\n" end # computer parsable report #def to_s # [ @antisense, @start, @stop ].join("\t") #end end # class Pair # = Bio::SiRNA::ShRNA # Designing shRNA. class ShRNA # Bio::Sequence::NA attr_accessor :top_strand # Bio::Sequence::NA attr_accessor :bottom_strand # Input is a Bio::SiRNA::Pair object (the target sequence). def initialize(pair) @pair = pair end # only the 'BLOCK-iT' rule is implemented for now. def design(method = 'BLOCK-iT') case method when 'BLOCK-iT' block_it else raise NotImplementedError end end # same as design('BLOCK-iT'). # method can be one of 'piGENE' (default) and 'BLOCK-iT'. def block_it(method = 'piGENE') top = Bio::Sequence::NA.new('CACC') # top_strand_shrna_overhang bot = Bio::Sequence::NA.new('AAAA') # bottom_strand_shrna_overhang fwd = @pair.sense rev = @pair.sense.complement case method when 'BLOCK-iT' # From BLOCK-iT's manual loop_fwd = Bio::Sequence::NA.new('CGAA') loop_rev = loop_fwd.complement when 'piGENE' # From piGENE document loop_fwd = Bio::Sequence::NA.new('GTGTGCTGTCC') loop_rev = loop_fwd.complement else raise NotImplementedError end if /^G/i =~ fwd @top_strand = top + fwd + loop_fwd + rev @bottom_strand = bot + fwd + loop_rev + rev else @top_strand = top + 'G' + fwd + loop_fwd + rev @bottom_strand = bot + fwd + loop_rev + rev + 'C' end end # human readable report def report report = "### shRNA\n" report << "Top strand shRNA (#{@top_strand.length} nt):\n" report << " 5'-#{@top_strand.upcase}-3'\n" report << "Bottom strand shRNA (#{@bottom_strand.length} nt):\n" report << " 3'-#{@bottom_strand.reverse.upcase}-5'\n" end end # class ShRNA end # class SiRNA end # module Bio if __FILE__ == $0 seq = Bio::Sequence::NA.new(ARGF.read) sirna = Bio::SiRNA.new(seq) pairs = sirna.design # or .design('uitei') or .uitei or .reynolds pairs.each do |pair| puts pair.report shrna = Bio::SiRNA::ShRNA.new(pair) shrna.design # or .design('BLOCK-iT') or .block_it puts shrna.report puts "# as DNA" puts shrna.top_strand.dna puts shrna.bottom_strand.dna end end =begin = ChangeLog 2005/03/21 Itoshi NIKAIDO Bio::SiRNA#ShRNA_designer method was changed design method. 2004/06/25 Bio::ShRNA class was added. 2004/06/17 Itoshi NIKAIDO We can use shRNA loop sequence from piGene document. =end