Extracellular Vesicle Delivery of Inhibitory Nucleic Acids for Bacterial Gene Modulation
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Solution Overview
Problem
Current methods are limited in their ability to functionally identify and manipulate bacterial genes within the human microbiota, particularly due to the inaccessibility of over 90% of intestinal bacteria to genetic manipulation techniques, hindering the understanding and treatment of diseases associated with microbiota changes.
Innovation Solution
The use of extracellular vesicles (EVs) derived from mammalian cells, loaded with inhibitory nucleic acids, to target and modulate bacterial gene expression in prokaryotic cells, enabling the regulation of specific genes associated with disease phenotypes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional genetic manipulation techniques (Flp-FRT recombinase, transposon mutagenesis, chemical screens) are used to identify and manipulate bacterial genes, then functional identification of bacterial genes can be achieved, but over 90% of intestinal bacteria remain inaccessible to these techniques
Solution Approach 1:
The patent uses extracellular vesicles (EVs) as an intermediary carrier to deliver inhibitory nucleic acids (siRNAs) to bacterial cells. The EVs are engineered to specifically target and fuse with bacterial membranes, enabling delivery of gene-silencing molecules to previously inaccessible bacterial species without requiring direct genetic manipulation tools
Solution Approach 2:
The patent replaces mechanical genetic manipulation methods (recombinase enzymes, transposons, chemical mutagens) with a biochemical approach using RNA interference. Instead of physically inserting or modifying bacterial DNA, the system uses siRNAs delivered via EVs to silence gene expression post-transcriptionally, bypassing the need for bacterial genetic manipulation machinery
2Adaptability or versatility
If extracellular vesicles are used to deliver inhibitory nucleic acids to bacterial cells, then gene expression modulation can be achieved in previously inaccessible bacteria, but the complexity of the delivery system increases
Solution Approach 1:
The patent creates a universal EV-based delivery platform that can target multiple different bacterial species using the same fundamental mechanism. The EVs are designed with universal features (membrane fusion capability, endosomal escape mechanism) while allowing customization of surface targets to reach different bacteria, reducing the need for species-specific delivery systems
Solution Approach 2:
The patent employs a nested structure where inhibitory nucleic acids are packaged inside extracellular vesicles, which themselves are derived from mammalian cells. The EVs contain the siRNA molecules protected within their lipid bilayer membrane, creating a protective nested architecture that shields the fragile nucleic acids during delivery through harsh biological environments
3Reliability
If inhibitory nucleic acids are delivered to prokaryotic cells using eukaryotic-derived extracellular vesicles, then bacterial gene expression can be modulated in vivo, but the processing and functioning of inhibitory nucleic acids requires mammalian machinery
Solution Approach 1:
The patent performs preliminary processing of the inhibitory nucleic acids within the eukaryotic mammalian cells that produce the EVs. The mammalian cellular machinery processes and matures the siRNAs before they are packaged into EVs and delivered to bacteria, ensuring the nucleic acids are in their active, processed form before encountering the prokaryotic target
Solution Approach 2:
The EVs act as an intermediary that bridges the eukaryotic processing machinery and the prokaryotic target cells. The EVs carry pre-processed mammalian machinery components and mature siRNAs across the species boundary, enabling the transfer of gene-silencing capability from eukaryotic to prokaryotic systems without requiring the bacteria to possess mammalian processing enzymes
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 allows for the effective modulation of bacterial gene expression in vivo, providing a potential therapeutic strategy for diseases such as metabolic disorders, inflammatory diseases, and cancers by modifying the microbiota composition.
Implementation Method 1
the one or more inhibitory nucleic acids target (e.g., hybridize to) one or more genes in the prokaryotic cell
Data Source
AI summary
Aspects of the disclosure relate to compositions and methods for delivering inhibitory nucleic acids to prokaryotic cells using extracellular vesicles (e.g., exosomes, microvesicles, etc.) derived from mammalian cells. The disclosure provides methods for regulating the expression of one or more genes in a prokaryotic cell. The disclosure further provides methods of treating diseases associated with prokaryotic gene dysregulation.