Correction to 'Influence of Fluorescent Protein Maturation on FRET Measurements in Living Cells'
| Autor: | Jonas van den Berg, Arnold J. Boersma, Liesbeth M. Veenhoff, Sebastian Kragh Kristensen, Bert Poolman, Sara N Mavrova, Luca Mantovanelli, Boqun Liu |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
macromolecular crowding
Green Fluorescent Proteins Bioengineering Saccharomyces cerevisiae FRET sensors Models Biological Maltose-Binding Proteins Article Cell Physiological Phenomena Escherichia coli Fluorescence Resonance Energy Transfer Fluorescent protein Promoter Regions Genetic Instrumentation Fluid Flow and Transfer Processes Protein Synthesis Inhibitors Microscopy Confocal Base Sequence Chemistry Process Chemistry and Technology fluorescent protein maturation biosensors Cell biology Addition/Correction Förster resonance energy transfer Chloramphenicol Gene Expression Regulation Microscopy Fluorescence Mutation |
| Zdroj: | ACS Sensors |
| ISSN: | 2379-3694 |
| Popis: | Förster resonance energy transfer (FRET)-based sensors are a valuable tool to quantify cell biology, yet it remains necessary to identify and prevent potential artifacts in order to exploit their full potential. We show here that artifacts arising from slow donor mCerulean3 maturation can be substantially diminished by constitutive expression in both prokaryotic and eukaryotic cells, which can also be achieved by incorporation of faster-maturing FRET donors. We developed an improved version of the donor mTurquoise2 that matures faster than the parent protein. Our analysis shows that using equal maturing fluorophores in FRET-based sensors or using constitutive low expression conditions helps to reduce maturation-induced artifacts, without the need of additional noise-inducing spectral corrections. In general, we show that monitoring and controlling the maturation of fluorescent proteins in living cells is important and should be addressed in in vivo applications of genetically encoded FRET sensors. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|