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Conditional fluorescent mouse translocation reporters for ERK1/2 and AKT signaling
Colin J Dinsmore1, Philippe Soriano1
1Department of Cell, Developmental, and Regenerative Biology, Icahn School of Medicine at Mt. Sinai, New York, NY, 10029, USA.
Abstract:
Understanding how cells activate intracellular signaling pathways in response to external signals, such as growth factors, is a longstanding goal of cell and developmental biology. Recently, live-cell signaling reporters have greatly expanded our understanding of signaling dynamics in response to wide-ranging stimuli and chemical or genetic perturbation, both ex vivo (cell lines) and in vivo (whole embryos or animals). Among the many varieties of reporter systems, translocation reporters that change sub-cellular localization in response to pathway activation have received considerable attention for their ease of use compared to FRET systems and favorable response times compared to transcriptional reporters. We reasoned that mouse reporter lines expressed in a conditional fashion would be a useful addition to the arsenal of mouse genetic tools, as such lines remain undeveloped despite widespread use of these sensors. We therefore created and validated two novel mouse reporter lines at the ROSA26 locus. One expresses an ERK1/2 pathway reporter and a nuclear marker from a single transcript, while the second additionally expresses an AKT reporter in order to simultaneously interrogate both pathways.
Insights
Researchers developed novel conditional mouse reporter lines to track cellular signaling pathways. These tools enable simultaneous monitoring of the ERK1/2 and AKT pathways, advancing cell biology research.
Area of Science:
- Cell Biology
- Developmental Biology
- Molecular Biology
Background:
- Understanding intracellular signaling pathways is crucial in cell and developmental biology.
- Live-cell signaling reporters, especially translocation reporters, offer advantages in studying signaling dynamics.
- Existing mouse genetic tools lack conditional reporter lines for simultaneous pathway analysis.
Purpose of the Study:
- To create and validate novel conditional mouse reporter lines for studying intracellular signaling.
- To enable simultaneous interrogation of the ERK1/2 and AKT pathways using a single genetic tool.
- To expand the available mouse genetic tools for live-cell signaling research.
Main Methods:
- Generation of two novel mouse reporter lines at the ROSA26 locus.
- Utilizing conditional expression for targeted reporter gene activation.
- Developing reporters for simultaneous monitoring of ERK1/2 and AKT pathway activity.
Main Results:
- Successfully created and validated two distinct conditional mouse reporter lines.
- One line reports on the ERK1/2 pathway and includes a nuclear marker.
- The second line reports on both ERK1/2 and AKT pathways simultaneously.
Conclusions:
- The developed mouse reporter lines are valuable additions to the toolkit for studying cell signaling.
- These reporters facilitate simultaneous, live-cell analysis of key signaling pathways in vivo.
- The conditional nature of these lines allows for precise temporal and spatial control of reporter expression.
