Repurposing a Two-Component System-Based Biosensor for the Killing of Vibrio cholerae.

Area of Science:

• Synthetic biology
• Microbiology
• Molecular biology

Background:

• Cholera remains a significant global health threat requiring novel control strategies.
• Current treatments for cholera are limited, necessitating innovative therapeutic approaches.
• Targeting the causative agent, Vibrio cholerae, is crucial for effective cholera management.

Purpose of the Study:

• To develop and validate an in vitro sense-and-kill system using engineered Escherichia coli to combat Vibrio cholerae.
• To create a synthetic biology-based approach for detecting and eliminating V. cholerae.
• To lay the groundwork for a potential probiotic treatment for cholera.

Main Methods:

• Engineered a wild-type Escherichia coli strain with a synthetic biology sense-and-kill system.
• Utilized V. cholerae's quorum-sensing molecule CAI-1 for specific pathogen detection.
• Incorporated lysis protein YebF-Art-085 and killing protein Art-085 for targeted V. cholerae elimination.
• Characterized protein expression and activity within a V. cholerae biosensing circuit.

Main Results:

• The engineered E. coli successfully detected V. cholerae via CAI-1.
• The system demonstrated self-lysis of E. coli to release Art-085 for V. cholerae killing.
• In the presence of V. cholerae supernatant, the integrated system effectively inhibited V. cholerae growth.
• Individual components YebF-Art-085 and Art-085 showed expected expression and activity.

Conclusions:

• The developed sense-and-kill system represents a proof-of-concept for targeting V. cholerae.
• This synthetic biology approach shows promise for a novel probiotic cholera treatment.
• Further development could lead to future preventative and therapeutic strategies for cholera.