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A Method to Discover Phased siRNA Loci

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Plant MicroRNAs

Part of the book series: Methods in Molecular Biology ((MIMB,volume 592))

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.

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Author information

Authors and Affiliations

  1. Department of Biology, Pennsylvania State University, University Park, PA, USA

    Michael J. Axtell

Authors
  1. Michael J. Axtell
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Editor information

Editors and Affiliations

  1. Delaware Biotechnology Institute, University of Delaware, Innovation Way 15, Newark, 19711, U.S.A.

    Blake C. Meyers

  2. Delaware Biotechnology Institute, University of Delaware, Innovation Way 15, Newark, 19711, U.S.A.

    Pamela J. Green

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© 2010 Humana Press, a part of Springer Science+Business Media, LLC

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Axtell, M.J. (2010). A Method to Discover Phased siRNA Loci. In: Meyers, B., Green, P. (eds) Plant MicroRNAs. Methods in Molecular Biology, vol 592. Humana Press. https://doi.org/10.1007/978-1-60327-005-2_5

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  • DOI: https://doi.org/10.1007/978-1-60327-005-2_5

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-004-5

  • Online ISBN: 978-1-60327-005-2

  • eBook Packages: Springer Protocols

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