Publication
Title
Fluorescent in situ hybridization for miRNA combined with staining of proteins
Author
Abstract
The last decades have illustrated the importance of microRNAs (miRNAs) in various biological and pathological processes. The combined visualization of miRNAs using fluorescent in situ hybridization (FISH) and proteins using immunofluorescence (IF) can reveal their spatiotemporal distribution in relation to the cell and tissue morphology and can provide interesting insights into miRNA‐protein interactions. However, standardized protocols for co‐localization of miRNAs and proteins are currently lacking, and substantial technical obstacles still need to be addressed. In particular, the incompatibility of protein IF protocols with steps required for miRNA FISH, such as proteolytic pretreatments and ethylcarbodiimide post‐fixation, as well as hurdles related to low signal intensity of low‐copy miRNAs, remains challenging. Our technique may considerably enhance miRNA‐based research, as current detection techniques lack the ability to elucidate cellular and subcellular localization. Here, we describe an optimized 2‐day protocol for combined detection of low‐abundant miRNAs and proteins in cryosections of cardiac tissue, without the need for protease‐dependent pretreatment or post‐fixation treatment. We successfully demonstrate endothelial‐specific localization of low‐abundant miR‐181c‐5p in cardiac tissue. © 2023 Wiley Periodicals LLC.
Language
English
Source (journal)
Current protocols
Publication
2023
DOI
10.1002/CPZ1.880
Volume/pages
3 :9 (2023)
Article Reference
e880
Pubmed ID
37728252
Medium
E-only publicatie
Full text (Publisher's DOI)
Full text (open access)
UAntwerpen
Faculty/Department
Research group
Publication type
Subject
Affiliation
Publications with a UAntwerp address
External links
Record
Identifier
Creation 26.10.2023
Last edited 25.04.2024
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