CRISPR-Protected Transducing Particles for Stable Bacterial Gene Delivery

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Solution Overview

Problem

There is a need for improved systems and methods to effectively deliver nucleic acid sequences of interest to target cells, particularly bacterial cells, and to manipulate bacterial populations, while addressing challenges in the production and safety of synthetic bacteriophages for therapeutic applications.

Innovation Solution

A system comprising a transducing particle with a CRISPR protective array that targets a selective component, a helper transducing particle, and a regulatory region to control CRISPR expression, enabling efficient delivery and protection of nucleic acid sequences to target cells, while ensuring safety and stability.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If synthetic bacteriophages are engineered to deliver nucleic acid sequences to target bacteria, then the therapeutic potential and precision of bacterial population manipulation is improved, but the complexity of production and safety control deteriorates

Engineering Contradiction:

Improveprecision of bacterial population manipulationVSAvoidcomplexity of production and safety control

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The system is divided into distinct functional modules: a transducing particle containing the nucleic acid sequence of interest, a helper particle providing essential functions, and a CRISPR-based selective component. This segmentation allows independent optimization of each module and simplifies production by separating complex functions into manageable components that can be produced and controlled separately.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The helper transducing particle acts as an intermediary that provides essential functions for the transducing particle without being the primary delivery vehicle. This intermediary approach allows the main transducing particle to focus on delivering the nucleic acid sequence while the helper particle handles complex production and safety functions, reducing overall system complexity.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If CRISPR protective array is used to target selective components, then the safety and stability of transducing particles is improved, but the device complexity increases

Engineering Contradiction:

Improvesafety and stability of transducing particlesVSAvoidcomplexity of CRISPR expression control

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The CRISPR protective array is pre-configured in the transducing particle to recognize and eliminate unwanted selective components before they can cause safety issues. This preliminary action ensures safety is built into the system from the outset rather than requiring complex post-production safety controls, improving reliability without proportionally increasing complexity.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The CRISPR system serves multiple functions: it provides safety by targeting selective components, maintains stability of the transducing particle population, and can be regulated by available bacterial regulatory regions. This multi-functionality reduces the need for separate safety mechanisms, balancing reliability improvement with controlled complexity.

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Stability of the object's composition

If regulatory regions are used to control CRISPR expression, then the stability of nucleic acid delivery is improved, but the manufacturing precision requirements increase

Engineering Contradiction:

Improvestability of nucleic acid deliveryVSAvoidprecision of CRISPR expression control

Core Design Contradiction:

Stability of the object's compositionVSManufacturing precision

Solution Approach 1:

The system uses regulatory regions that are naturally recognized by the target bacterial host's own transcriptional machinery to control CRISPR expression. This self-service approach leverages the host's existing regulatory pathways rather than requiring externally imposed control mechanisms, improving stability while reducing manufacturing precision requirements since the host cell itself performs the regulation.

Inventive Principle:

Principle #25Self-service

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

The system enables precise delivery and manipulation of nucleic acid sequences in bacterial populations, creating a selection pressure that favors cells carrying the transduced sequence, enhancing therapeutic potential and safety.

Implementation Method 1

at least one spacer of the CRISPR protective array targets at least one proto-spacer comprised within at least one selective component, so as to specifically inactivate the selective component

Methodology Applied

Scientific EffectCRISPR array targeting:

Data Source

PatentUS20260083787A1Systems for production of transducing particles, methods, kits, compositions and uses thereof

Publication Date: 2026.03.26 TROBIX BIO LTD

AI summary

The present disclosure provides improved systems and methods for production of bacteriophage-based engineered transducing particles and uses thereof in manipulating bacterial populations to express any nucleic acid sequence of interest. The disclosed systems are based on inserting to producing cells nucleic acid molecules that comprise the nucleic acid sequence of interest and a protective regulated array, and regulators specific for the regulator array. The system further provides helper transducing particle that facilitates the propagation of the transducing particle.