Programmable trans-splicing riboregulators for complex cellular logic computation

Area of Science:

• Synthetic biology
• Molecular biology
• Biotechnology

Background:

• Synthetic genetic circuits require reliable regulatory mechanisms for scalability.
• Existing systems face limitations in programmability, performance, predictability, and orthogonality.

Purpose of the Study:

• To develop a novel class of regulatory elements for synthetic genetic circuits.
• To enhance the programmability, predictability, and orthogonality of gene regulation.

Main Methods:

• Design and implementation of split-intron-enabled trans-splicing riboregulators (SENTRs).
• Utilizing de novo designed external guide sequences for riboregulator function.
• Employing machine learning for predictability assessment.

Main Results:

• SENTRs exhibit low leakage expression, wide dynamic range, and low crosstalk.
• Demonstrated RNA sensing, logic computation, and signal transduction capabilities.
• Successfully implemented digital logic operations with up to six inputs using orthogonal SENTRs.

Conclusions:

• SENTRs provide a powerful and versatile post-transcriptional regulatory tool.
• The technology enables complex gene circuit construction and offers broad biotechnological applications.
• SENTRs advance the field of synthetic biology with improved genetic control.