Gene editing tool-loaded biomimetic cationic vesicles with highly efficient bacterial internalization for in vivo...

Area of Science:

• Microbiology
• Biotechnology
• Infectious Diseases

Background:

• Drug-resistant bacterial infections, including multidrug-resistant Acinetobacter baumannii (MRAB) and drug-resistant Pseudomonas aeruginosa (DRPA), are a major cause of death post-COVID-19.
• Classical antibiotic strategies are limited by the growing challenge of bacterial drug resistance.

Purpose of the Study:

• To develop a novel antimicrobial strategy using CRISPR/Cas9 plasmid delivery to combat drug-resistant bacterial infections.
• To overcome the challenge of delivering large, negatively charged CRISPR/Cas9 plasmids to pathogens at infection sites.

Main Methods:

• Construction of CRISPR/Cas9 plasmids designed to replicate within MRAB or DRPA and induce chromosomal damage.
• Development of biomimetic cationic hybrid vesicles (BCVs) integrating bacterial outer membrane vesicles and cationic lipids for efficient plasmid delivery.
• In vitro and in vivo testing of BCV-mediated plasmid delivery and subsequent eradication of MRAB and DRPA infections.

Main Results:

• BCVs demonstrated efficient in vitro and in vivo delivery and internalization of CRISPR/Cas9 plasmids into MRAB and DRPA.
• Intratracheal or topical hydrogel application of BCVs successfully eradicated MRAB pulmonary infection and DRPA wound infection.
• The CRISPR/Cas9 plasmid-loaded BCVs proved effective in eliminating target pathogens from infected lungs and wounds.

Conclusions:

• CRISPR/Cas9 plasmid-loaded BCVs represent a promising, resistance-independent therapeutic approach for treating drug-resistant bacterial infections.
• This novel delivery system overcomes previous limitations in delivering large genetic payloads to bacterial pathogens.
• The strategy shows potential for treating serious infections caused by multidrug-resistant Acinetobacter baumannii and drug-resistant Pseudomonas aeruginosa.