Measuring transcription dynamics in living cells using a photobleaching approach
- PMID: 28434903
- DOI: 10.1016/j.ymeth.2017.04.007
Measuring transcription dynamics in living cells using a photobleaching approach
Abstract
The transcriptional kinetics of RNA polymerase II, the enzyme responsible for mRNA transcription in the nucleoplasm, can be modulated by a variety of factors. It is therefore important to establish experimental systems that will enable the readout of transcription kinetics of specific genes as they occur in real time within individual cells. This can be performed by implementing fluorescent tagging of the mRNA under live-cell conditions. This chapter describes how to generate fluorescently tagged genes and mRNA, and how a photobleaching approach can produce information on mRNA transcription kinetics.
Copyright © 2017 Elsevier Inc. All rights reserved.
Similar articles
-
Fluorescence recovery after photobleaching.Methods Mol Biol. 2011;769:387-402. doi: 10.1007/978-1-61779-207-6_26. Methods Mol Biol. 2011. PMID: 21748690
-
Monitoring the Mitochondrial Dynamics in Mammalian Cells.Methods Mol Biol. 2018;1782:267-285. doi: 10.1007/978-1-4939-7831-1_15. Methods Mol Biol. 2018. PMID: 29851005
-
Measuring the kinetics of mRNA transcription in single living cells.J Vis Exp. 2011 Aug 25;(54):e2898. doi: 10.3791/2898. J Vis Exp. 2011. PMID: 21904295 Free PMC article.
-
Dynamics of mammalian NER proteins.DNA Repair (Amst). 2011 Jul 15;10(7):760-71. doi: 10.1016/j.dnarep.2011.04.015. Epub 2011 May 7. DNA Repair (Amst). 2011. PMID: 21550320 Review.
-
Real-time imaging of the HIV-1 transcription cycle in single living cells.Methods. 2011 Jan;53(1):62-7. doi: 10.1016/j.ymeth.2010.06.015. Epub 2010 Jun 25. Methods. 2011. PMID: 20600934 Review.
Cited by
-
Uncoupling of nucleo-cytoplasmic RNA export and localization during stress.Nucleic Acids Res. 2019 May 21;47(9):4778-4797. doi: 10.1093/nar/gkz168. Nucleic Acids Res. 2019. PMID: 30864659 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials