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Detection of MicroRNA Processing Intermediates Through RNA Ligation Approaches

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Plant MicroRNAs

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1932))

Abstract

MicroRNAs (miRNA) are small RNAs of 20–22 nt that regulate diverse biological pathways through the modulation of gene expression. miRNAs recognize target RNAs by base complementarity and guide them to degradation or translational arrest. They are transcribed as longer precursors with extensive secondary structures. In plants, these precursors are processed by a complex harboring DICER-LIKE1 (DCL1), which cuts on the precursor stem region to release the mature miRNA together with the miRNA*. In both plants and animals, the miRNA precursors contain spatial clues that determine the position of the miRNA along their sequences. DCL1 is assisted by several proteins, such as the double-stranded RNA binding protein, HYPONASTIC LEAVES1 (HYL1), and the zinc finger protein SERRATE (SE). The precise biogenesis of miRNAs is of utter importance since it determines the exact nucleotide sequence of the mature small RNAs and therefore the identity of the target genes. miRNA processing itself can be regulated and therefore can determine the final small RNA levels and activity. Here, we describe methods to analyze miRNA processing intermediates in plants. These approaches can be used in wild-type or mutant plants, as well as in plants grown under different conditions, allowing a molecular characterization of the miRNA biogenesis from the RNA precursor perspective.

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Acknowledgments

Supported by grants of Argentinean Ministry of Science, PICT-2015-3557 and PICT-2016-0761 to J.P. CONICET fellowships to B.M. and A.M.L.R.

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Authors and Affiliations

  1. Instituto de Biología Molecular y Celular de Rosario, CONICET, Universidad Nacional de Rosario, Rosario, Argentina

    Belén Moro, Arantxa M. L. Rojas & Javier F. Palatnik

  2. Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina

    Belén Moro

  3. Centro de Estudios Interdisciplinarios, Universidad Nacional de Rosario, Rosario, Argentina

    Javier F. Palatnik

Authors
  1. Belén Moro
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  2. Arantxa M. L. Rojas
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  3. Javier F. Palatnik
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Corresponding authors

Correspondence to Belén Moro or Javier F. Palatnik .

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Editors and Affiliations

  1. Unidad de Genómica Avanzada, Laboratorio Nacional de Genómica para la Biodiversidad (LANGEBIO), Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Irapuato, Guanajuato, Mexico

    Stefan de Folter

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Moro, B., Rojas, A.M.L., Palatnik, J.F. (2019). Detection of MicroRNA Processing Intermediates Through RNA Ligation Approaches. In: de Folter, S. (eds) Plant MicroRNAs. Methods in Molecular Biology, vol 1932. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9042-9_20

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  • DOI: https://doi.org/10.1007/978-1-4939-9042-9_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9041-2

  • Online ISBN: 978-1-4939-9042-9

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