Editor
Chiara Ambrogio
  • Faculty, Department of Molecular Biotechnology and Health Sciences, University of Torino
Research fields
  • Cancer Biology, Cell Biology, Molecular Biology
Biotinylated Micro-RNA Pull Down Assay for Identifying miRNA Targets
Authors:  Pornima Phatak and James M Donahue, date: 05/05/2017, view: 18427, Q&A: 1
microRNA (miRNA) directly associates with its target transcripts (mRNA). This protocol describes a method for detection of direct interaction between miRNA and mRNA. The result of interaction helps screening the specific target mRNAs for a miRNA.
In situ Hybridization (ISH) and Quantum Dots (QD) of miRNAs
Authors:  Sajni Josson, Murali Gururajan and Leland W.K. Chung, date: 02/20/2017, view: 10220, Q&A: 0
miRNA are short non-coding RNA which inhibit translation of mRNA. miRNA regulate several cellular processes. Certain miRNA are known to induce oncogenesis. miRNA can be measured by real-time PCR and be imaged using a combination of in situ hybridization (ISH) and quantum dots (QD). The advantage of using quantum dots is that several miRNA can be simultaneously measured using multiplexed QD. Additionally, miRNA can be visualized in different regions of the tissue. Since miRNA are biomarkers of various disease states, miRNA can be visualized and quantitated in tissue sections for diagnostic and prognostic purposes. Here we describe ISH-QD analysis of tissue sections. Tissue sections from xenografts or clinical specimens are used. These are deparaffinized, treated with Proteinase K and hybridized with a biotin-probe to specific to the miRNA. The in situ hybridization is performed by labeling the biotin-probes and followed by labeling with streptavidin tagged quantum dots. Image acquisition of the quantum dots is performed and analyzed for the miRNA expression levels. Combining ISH and QD gives a powerful tool to detect miRNA in different cells of the tissue.
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