Intercellular CRISPRi for Distributed Genetic Circuits.

Area of Science:

• Synthetic Biology
• Microbial Engineering
• Genetic Circuits

Background:

• Multicellular organisms and microbes distribute tasks among specialized cells for efficient resource allocation.
• Synthetic multicellular circuits distribute large genetic programs across multiple cells to reduce individual cell burden.

Purpose of the Study:

• To implement multicellular logic-gate circuits in bacterial co-cultures.
• To combine DNA messaging with CRISPR interference for programmable gene regulation.

Main Methods:

• Engineered sender bacteria to transmit DNA messages encoding guide RNAs.
• Receiver bacteria were designed to receive DNA messages and express guide RNAs.
• CRISPR interference (CRISPRi) was utilized to regulate transcription in receiver cells.

Main Results:

• Demonstrated functional multicellular circuits representing digital logic gates: NOT, YES, AND, and AND-AND-NOT.
• Circuits operated on timescales comparable to small molecule signaling.
• Receiver cells successfully processed inputs and generated logical outputs.

Conclusions:

• This work presents a novel system for building synthetic multicellular computational circuits.
• The DNA messaging and CRISPRi approach offers a programmable platform for complex cellular functions.
• The developed logic gates provide a foundation for more sophisticated synthetic biological systems.