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Detection of MicroRNA Processing Intermediates Through RNA Ligation Approaches.
Moro, Belén; Rojas, Arantxa M L; Palatnik, Javier F.
Afiliação
  • Moro B; Instituto de Biología Molecular y Celular de Rosario, CONICET, Universidad Nacional de Rosario, Rosario, Argentina. moro@ibr-conicet.gov.ar.
  • Rojas AML; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina. moro@ibr-conicet.gov.ar.
  • Palatnik JF; Instituto de Biología Molecular y Celular de Rosario, CONICET, Universidad Nacional de Rosario, Rosario, Argentina.
Methods Mol Biol ; 1932: 261-283, 2019.
Article em En | MEDLINE | ID: mdl-30701507
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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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Arabidopsis / RNA de Plantas / Proteínas de Arabidopsis / MicroRNAs Tipo de estudo: Diagnostic_studies Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Argentina País de publicação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Arabidopsis / RNA de Plantas / Proteínas de Arabidopsis / MicroRNAs Tipo de estudo: Diagnostic_studies Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Argentina País de publicação: Estados Unidos