Recombinant Phasmid CRISPR Delivery for Targeted Bacterial Eradication
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Solution Overview
Problem
Current antimicrobial therapies face challenges due to antibiotic resistance and disruptions in the gastrointestinal microbiota, leading to long-lasting metabolic changes and the emergence of resistant strains, with limited success in identifying new antibiotics that can effectively penetrate bacterial cell walls.
Innovation Solution
Development of recombinant host bacteria comprising a recombinant phasmid with a bacteriophage genome lacking replication and lysis modules, engineered to deliver CRISPR RNAs that target specific bacterial strains, altering the microbiome and treating infections by administering therapeutically effective amounts of these bacteria.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If antibiotics are used to eradicate bacterial pathogens, then bacterial infections are treated, but the gastrointestinal microbiota is disrupted leading to long-lasting metabolic changes and antibiotic resistance
Solution Approach 1:
The invention applies local quality by designing CRISPR-Cas systems with spacer sequences that are specific to particular bacterial pathogens. This allows the antimicrobial activity to be localized to only the target pathogen while leaving the rest of the microbiota untouched. The guide RNA is engineered to match only the genetic sequence of the specific pathogen, ensuring selective killing without broad-spectrum disruption.
Solution Approach 2:
The invention segments the antimicrobial function by separating the CRISPR-Cas system into modular components that can be independently designed and assembled. The spacer sequence is specifically tailored to match the target pathogen's genome, while the Cas protein and other components remain generic. This segmentation allows for pathogen-specific targeting without affecting other bacteria in the microbiota.
2Reliability
If traditional antibiotics are used, then bacterial infections are treated, but new antibiotics have limited success in penetrating bacterial cell walls
Solution Approach 1:
The invention uses an intermediary approach by employing bacteriophage-derived delivery vehicles to transport the CRISPR-Cas system into the target bacteria. These phage-based vectors naturally penetrate bacterial cell walls through their established infection mechanisms, solving the delivery problem without requiring the antimicrobial agent itself to have penetration capabilities. The CRISPR components are delivered as genetic material that is then expressed inside the target cell.
Solution Approach 2:
The invention replaces the mechanical approach of small-molecule antibiotic penetration with a biological delivery system. Instead of relying on chemical properties to penetrate the cell wall, the CRISPR-Cas system is delivered via bacteriophage vectors that use their natural infection machinery to inject genetic material into the bacterial cell, bypassing the cell wall barrier entirely.
3Reliability
If CRISPR-Cas systems are delivered to target bacteria, then specific bacterial strains are killed, but the complexity of delivering and expressing the system increases
Solution Approach 1:
The invention applies universality by using a standardized Cas protein and delivery vector platform that can be applied to multiple different pathogen targets. The core CRISPR-Cas machinery remains the same, but only the spacer sequence needs to be changed to target different bacteria. This multi-functional platform reduces overall system complexity by reusing proven components across different applications.
Solution Approach 2:
The invention uses preliminary action by pre-assembling the CRISPR-Cas system within bacteriophage vectors before delivery. The guide RNA and Cas protein are packaged into the phage particles in advance, ready for immediate expression upon infection. This pre-preparation simplifies the delivery process and ensures that all necessary components are present and functional before encountering the target pathogen.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach effectively kills target bacterial strains and alters the microbial population, providing a novel method to treat bacterial infections while avoiding common drug resistance mechanisms and minimizing disruptions to the microbiota.
Implementation Method 1
the spacer is at least about 70% complementary to a nucleic acid of the target bacterium
Data Source
AI summary
The invention is directed to antibacterial compositions comprising bacteria modified to comprise phasmids engineered to deliver of CRISPR RNAs and methods for their use.