Genetically-encoded tools for cAMP probing and modulation in living systems.

Area of Science:

• Cellular and Molecular Biology
• Biochemistry
• Synthetic Biology

Background:

• Intracellular 3'-5'-cyclic adenosine monophosphate (cAMP) is a critical second messenger in numerous signaling pathways, including those initiated by G protein-coupled receptors.
• Conventional biochemical assays for cAMP are limited by the need for sample disruption, low spatial resolution, and inability to perform kinetic measurements.
• These limitations hinder detailed understanding of cAMP dynamics and interactions within living cells.

Purpose of the Study:

• To review modern genetically-encoded tools for probing and modulating intracellular cAMP in living systems.
• To compare the capabilities and limitations of these novel tools.
• To provide guidance for selecting appropriate tools for specific experimental needs.

Main Methods:

• Discussion of genetically-encoded biosensors for cAMP detection.
• Overview of genetically-encoded actuators for cAMP modulation.
• Analysis of tool performance in various live-cell experimental setups.

Main Results:

• Genetically-encoded tools offer significant advantages over conventional assays, enabling real-time, spatially resolved measurements of cAMP.
• These tools allow for precise control over intracellular cAMP levels and dynamics.
• The review categorizes and evaluates different classes of tools based on their operational characteristics.

Conclusions:

• Modern genetically-encoded tools represent a significant advancement for studying intracellular cAMP.
• These tools overcome the limitations of traditional biochemical assays, facilitating deeper insights into cellular signaling.
• Rational selection of these tools is crucial for advancing research in cell physiology and drug discovery.