Programming biological communication between distinct membraneless compartments.

Area of Science:

• Synthetic biology
• Cell biology
• Biochemistry

Background:

• Membraneless organelles are crucial for cellular functions but creating synthetic communicating ones remains a challenge.
• Understanding the principles of membraneless organelle crosstalk is essential for designing complex cellular systems.

Purpose of the Study:

• To engineer a binary population of synthetic membraneless compartments capable of communication and feedback.
• To demonstrate programmable control over the assembly and function of these synthetic compartments.

Main Methods:

• Utilized two orthogonally phase-separating proteins in a cell-free expression system to form compartment consortia.
• Programmed the temporal and ordered appearance of compartments for controlled molecular delivery.
• Implemented protease and DNA-based molecular messages to trigger sensing, processing, and delivery of protein cargo.
• Integrated a feedback loop at the messenger RNA level to control information flow.

Main Results:

• Successfully created coexisting membraneless compartments with biological communication and controllable feedback.
• Demonstrated programmable, on-demand delivery of functional protein cargo in response to molecular cues.
• Showcased DNA-based programming for sensing and processing information, including a feedback mechanism.
• Established a design principle for constructing functional compartment consortia.

Conclusions:

• The engineered compartment consortia represent a novel approach to creating biosynthetic communicating membraneless organelles.
• These findings offer insights into the crosstalk mechanisms of natural membraneless organelles.
• Provides a foundational design principle for building complex, functional synthetic cellular systems.