Turn-on protein switches for controlling actin binding in cells

Area of Science:

• Cell Biology
• Biochemistry
• Synthetic Biology

Background:

• Filamentous actin (F-actin) is crucial for cellular functions, organized by numerous actin-binding proteins (ABPs).
• Studying individual ABPs and subcellularly controlling actin-binding is challenging.
• Harnessing the F-actin cytoskeleton for synthetic biology requires novel manipulation tools.

Purpose of the Study:

• To develop a novel system for externally controlling protein-actin interactions.
• To enable localized and stimulus-responsive manipulation of the F-actin cytoskeleton.
• To explore applications in synthetic biology for controlling cellular and tissue behavior.

Main Methods:

• Design and engineering of Controllable Actin-binding Switch Tools (CASTs).
• Development of CASTs responsive to various external stimuli with tunable kinetics.
• Genetic encoding of CASTs for insertion into native proteins and engineered structures.

Main Results:

• Demonstrated externally controllable actin-binding behavior of CASTs.
• Achieved orthogonal and multiplexed control over F-actin association.
• Showcased localized control of F-actin by inserting CASTs into native proteins.
• Engineered CASTs into structures to influence cell and tissue shape and behavior.

Conclusions:

• CASTs provide a powerful, genetically encoded platform for precise, stimulus-responsive control of F-actin.
• This technology overcomes limitations in studying ABPs and manipulating the actin cytoskeleton.
• CASTs open new avenues for synthetic biology, cell engineering, and understanding F-actin dynamics.