Dual Fluorescence Reporter Based Analytical Flow Cytometry for miRNA Induced Regulation in Mammalian Cells
Autor: | Liezel Tamon, Nicolas Lemus-Diaz, Jens Gruber |
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Jazyk: | angličtina |
Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Regulation of gene expression education.field_of_study Small RNA medicine.diagnostic_test Strategy and Management Mechanical Engineering Systems biology Population Metals and Alloys Computational biology Biology Non-coding RNA Industrial and Manufacturing Engineering Flow cytometry 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Single-cell analysis RNA interference 030220 oncology & carcinogenesis medicine Methods Article education |
Zdroj: | Bio Protoc |
Popis: | 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. |
Databáze: | OpenAIRE |
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