Localization of miRNAs by In Situ Hybridization in Plants Using Conventional Oligonucleotide Probes. 2017

Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
Unidad de Biotecnología, Centro de Investigación Científica de Yucatán, Calle 43 No. 130, Col. Chuburná de Hidalgo, Merida, Yucatán, 97200, Mexico.

Among the epigenetic mechanisms studied with a greater interest in the last decade are the microRNAs (miRNAs). These small noncoding RNA sequences that are approximately 17-22 nucleotides in length play an essential role in many biological processes of various organisms, including plants. The analysis of spatiotemporal expression of miRNAs provides a better understanding of the role of these small molecules in plant development, cell differentiation, and other processes; but such analysis is also an important method for the validation of biological functions. In this work, we describe the optimization of an efficient protocol for the spatiotemporal analysis of miRNA by in situ hybridization using different plant tissues embedded in paraffin. Instead of LNA-modified probes that are typically used for this work, we use conventional oligonucleotide probes that yield a high specificity and clean distribution of miRNAs.

UI MeSH Term Description Entries
D007150 Immunohistochemistry Histochemical localization of immunoreactive substances using labeled antibodies as reagents. Immunocytochemistry,Immunogold Techniques,Immunogold-Silver Techniques,Immunohistocytochemistry,Immunolabeling Techniques,Immunogold Technics,Immunogold-Silver Technics,Immunolabeling Technics,Immunogold Silver Technics,Immunogold Silver Techniques,Immunogold Technic,Immunogold Technique,Immunogold-Silver Technic,Immunogold-Silver Technique,Immunolabeling Technic,Immunolabeling Technique,Technic, Immunogold,Technic, Immunogold-Silver,Technic, Immunolabeling,Technics, Immunogold,Technics, Immunogold-Silver,Technics, Immunolabeling,Technique, Immunogold,Technique, Immunogold-Silver,Technique, Immunolabeling,Techniques, Immunogold,Techniques, Immunogold-Silver,Techniques, Immunolabeling
D015345 Oligonucleotide Probes Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin. Oligodeoxyribonucleotide Probes,Oligonucleotide Probe,Oligoribonucleotide Probes,Probe, Oligonucleotide,Probes, Oligodeoxyribonucleotide,Probes, Oligonucleotide,Probes, Oligoribonucleotide
D016612 Paraffin Embedding The infiltrating of tissue specimens with paraffin, as a supporting substance, to prepare for sectioning with a microtome. Embedding, Paraffin
D016707 Tissue Fixation The technique of using FIXATIVES in the preparation of cytologic, histologic, or pathologic specimens for the purpose of maintaining the existing form and structure of all the constituent elements. Fixation, Tissue
D017403 In Situ Hybridization A technique that localizes specific nucleic acid sequences within intact chromosomes, eukaryotic cells, or bacterial cells through the use of specific nucleic acid-labeled probes. Hybridization in Situ,Hybridization, In Situ,Hybridizations, In Situ,In Situ Hybridizations
D018749 RNA, Plant Ribonucleic acid in plants having regulatory and catalytic roles as well as involvement in protein synthesis. Plant RNA
D035683 MicroRNAs Small double-stranded, non-protein coding RNAs, 21-25 nucleotides in length generated from single-stranded microRNA gene transcripts by the same RIBONUCLEASE III, Dicer, that produces small interfering RNAs (RNA, SMALL INTERFERING). They become part of the RNA-INDUCED SILENCING COMPLEX and repress the translation (TRANSLATION, GENETIC) of target RNA by binding to homologous 3'UTR region as an imperfect match. The small temporal RNAs (stRNAs), let-7 and lin-4, from C. elegans, are the first 2 miRNAs discovered, and are from a class of miRNAs involved in developmental timing. RNA, Small Temporal,Small Temporal RNA,miRNA,stRNA,Micro RNA,MicroRNA,Primary MicroRNA,Primary miRNA,miRNAs,pre-miRNA,pri-miRNA,MicroRNA, Primary,RNA, Micro,Temporal RNA, Small,miRNA, Primary,pre miRNA,pri miRNA

Related Publications

Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
November 1990, Genes, chromosomes & cancer,
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
January 2020, Methods in molecular biology (Clifton, N.J.),
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
January 2002, International review of neurobiology,
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
December 1990, Journal of cellular biochemistry,
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
January 2006, Nature methods,
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
January 1985, Annals of the New York Academy of Sciences,
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
February 2012, Nature protocols,
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
May 1996, Applied and environmental microbiology,
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
January 2007, Advances in clinical chemistry,
Sara Hernández-Castellano, and Geovanny I Nic-Can, and Clelia De-la-Peña
October 2007, Methods (San Diego, Calif.),
Copied contents to your clipboard!