NDLI: Dual Fluorescence Reporter Based Analytical Flow Cytometry for miRNA Induced Regulation in Mammalian Cells.

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Dual Fluorescence Reporter Based Analytical Flow Cytometry for miRNA Induced Regulation in Mammalian Cells.

Content Provider	Europe PMC
Author	Lemus-Diaz, Nicolas Tamon, Liezel Gruber, Jens
Copyright Year	2018
Abstract	MicroRNA-induced gene regulation is a growing field in basic and translational research. Examining this regulation directly in cells is necessary to validate high-throughput data originated from RNA sequencing technologies. For this several studies employ luciferase-based reporters that usually measure the whole cell population, which comes with low resolution for the complexity of the miRNA-induced regulation. Here, we provide a protocol using a dual-fluorescence reporter and flow cytometry reaching single cell resolution; the protocol contains a simplified workflow that includes: vector generation, data acquisition, processing, and analysis using the R environment. Our protocol enables high-resolution measurements of miRNA induced post-transcriptional gene regulation and combined with system biology it can be used to estimate miRNAs proficiency.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC8328604&blobtype=pdf
Volume Number	8
PubMed Central reference number	PMC8328604
Issue Number	17
PubMed reference number	34395794
Journal	Bio-protocol [Bio Protoc]
e-ISSN	23318325
DOI	10.21769/bioprotoc.3000
Language	English
Publisher	Bio-Protocol
Publisher Date	2018-09-05
Publisher Place	1075 Lorne Way, Sunnyvale, CA 94087, USA
Access Restriction	Open
Rights License	Copyright © 2018 The Authors; exclusive licensee Bio-protocol LLC.
Subject Keyword	miRNA Flow-Cytometry Dual fluorescence reporter Functional assay ncRNA Small RNA non-protein-coding RNA Gene regulation Reporter gene system Single cell analysis Flow cytometry
Content Type	Text
Resource Type	Article
Subject	Immunology and Microbiology Neuroscience Plant Science Biochemistry, Genetics and Molecular Biology

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