Dual Fluorescence Reporter Based Analytical Flow Cytometry for miRNA Induced Regulation in Mammalian Cells
| Autor: | Nicolas Lemus-Diaz, Liezel Tamon, Jens Gruber |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: | |
| Zdroj: | Bio-Protocol, Vol 8, Iss 17 (2018) |
| Druh dokumentu: | article |
| ISSN: | 2331-8325 23180447 |
| DOI: | 10.21769/BioProtoc.3000 |
| 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: | Directory of Open Access Journals |
| Externí odkaz: |
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