Hierarchical control of enzymatic actuators using DNA-based switchable memories

Area of Science:

• Biochemistry
• Molecular Biology
• Synthetic Biology

Background:

• Cellular signaling networks inspire DNA-based programming for cell-free biochemical systems.
• Coupling DNA-based networks to downstream processes remains a significant challenge in molecular programming.

Purpose of the Study:

• To engineer hierarchical control of enzymatic actuators using feedback-controlled DNA circuits.
• To expand the capabilities of molecular programming for advanced regulatory and computational functions.

Main Methods:

• Developed a translator module to convert signaling molecules into DNA strands for actuator control.
• Engineered feedback-controlled DNA circuits for hierarchical regulation.
• Coupled DNA circuits to downstream enzymatic actuators (β-lactamase and luciferase).
• Performed detailed computational analysis to support findings.

Main Results:

• Successfully engineered hierarchical control of enzymatic actuators via DNA circuits.
• Demonstrated minimal retroactivity in the translator module.
• Showcased a modular approach by coupling a switchable memories circuit to downstream actuators.
• Achieved advanced regulatory dynamics in a cell-free system.

Conclusions:

• This work presents one of the most complex and versatile DNA-based circuits to date.
• The modular approach facilitates the engineering of sophisticated, hierarchical biochemical networks.
• This advancement broadens the scope of molecular programming for diverse applications.