A method to discover phased siRNA loci

Methods Mol Biol. 2010:592:59-70. doi: 10.1007/978-1-60327-005-2_5.

Abstract

Short, interfering RNAs (siRNAs) arise from the processing of long double-stranded RNA (dsRNA) by Dicer enzymes. Dicers generate siRNA duplexes by successive hydrolysis of both strands of the dsRNA phosphodiester backbone at positions determined by measuring 21-24 nucleotides from an exposed dsRNA terminus. Therefore, a population of dsRNAs with precisely identical termini will produce siRNA spaced in regular, 21-24-nucleotide intervals. This chapter presents an easily customized and generally applicable strategy for identifying loci which produce the "phased" siRNAs diagnostic of such processing. Given the input of a large set of expressed small RNAs and of the corresponding genome or transcriptome from which the small RNAs are derived, the methodology produces a ranking of user-defined loci with respect to their likely production of phased siRNAs. Top ranked loci are candidates for further computational and biological analyses.

MeSH terms

  • Computational Biology / methods*
  • RNA, Plant / genetics
  • RNA, Small Interfering / genetics*

Substances

  • RNA, Plant
  • RNA, Small Interfering