Contractile Injection Cargo Loading for Eukaryotic Cell Delivery
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Solution Overview
Problem
The range of hosts targeted by Contractile Injection Systems (CIS) like Metamorphosis Associated Contractile structures (MACs) and the identity and mode of action of effectors delivered to target cells in eukaryotes remain poorly understood, limiting their application in delivering proteinaceous cargos effectively.
Innovation Solution
A chimeric product comprising a recombinant bacterial Contractile Injection System (CIS) or Metamorphosis Associated Contractile structure (MAC) with a Metamorphosis-Inducing Factor 1 (Mif1) protein and a chaperone 605 protein, associated with a proteinaceous cargo, is used to deliver proteins, peptides, or compounds into eukaryotic cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a recombinant bacterial Contractile Injection System (CIS) or Metamorphosis Associated Contractile structure (MAC) is used to deliver effectors to target cells, then the delivery capability to eukaryotic cells is improved, but the understanding of host range and effector mode of action remains poorly understood, limiting effective application
Solution Approach 1:
The patent uses Mif1 protein as an intermediary effector that can be loaded into the CIS/MAC system. This intermediary allows researchers to study the delivery mechanism systematically, as Mif1's function in inducing metamorphosis provides a measurable readout for understanding how CIS/MAC systems interact with eukaryotic cells and what determines host range.
Solution Approach 2:
The patent segments the CIS/MAC system into distinct components (the injection system itself, the Mif1 effector protein, and the chaperone 605 protein) that can be independently studied and recombined. This segmentation allows systematic investigation of each component's role in delivery, host recognition, and effector function, thereby addressing the knowledge gaps about host range and mode of action.
2Stability of the object's composition
If the identity and mode of action of effectors delivered to target cells remain poorly understood, then the basic biological interactions are maintained, but the application in delivering proteinaceous cargos effectively is limited
Solution Approach 1:
The patent creates a universal delivery platform by combining the CIS/MAC system with Mif1 and chaperone 605, which can deliver various proteinaceous cargos. The Mif1-chaperone 605 complex serves as a versatile module that can be adapted to deliver different effectors, thereby enabling effective application across multiple contexts while maintaining the core biological interaction mechanism.
Solution Approach 2:
The patent changes the parameters of the effector delivery system by introducing Mif1 with specific sequence identity thresholds (at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100% sequence identity to SEQ ID NO:1) and defining specific functional properties (inducing metamorphosis). These parameter definitions transform the poorly understood effector delivery into a controllable, measurable process that can be effectively applied.
3Adaptability or versatility
If chimeric products comprising CIS/MAC with Mif1 and chaperone 605 are constructed, then targeted delivery of proteinaceous cargos is enabled, but the complexity of the system increases
Solution Approach 1:
The patent employs a nested structure where Mif1 protein is positioned in the inner core of the CIS/MAC tube, chaperone 605 is associated with Mif1, and the entire complex is delivered by the CIS/MAC system into the target cell. This nested organization (CIS/MAC containing Mif1 containing cargo) enables targeted delivery while systematically managing complexity through hierarchical structuring.
Solution Approach 2:
The patent merges multiple functional elements (CIS/MAC injection system, Mif1 effector protein, chaperone 605) into a single chimeric product that functions as an integrated delivery vehicle. This merging creates a versatile targeted delivery system where the combined components work together synergistically, and the system can be manufactured as a unified product rather than separate elements.
Data Source
AI summary
Provided are compositions, kits, and methods for delivering a proteinaceous cargo, or a protein or a peptide, or a drug or a marker, to or into a cell or to an individual in need thereof. In alternative embodiments, products of manufacture as provided herein comprise: (a) a recombinant bacterial Contractile Injection System (CIS) or a Metamorphosis Associated Contractile structure (MAC) formed or configured to comprise a tube having an inner core, (b) a Metamorphosis-Inducing Factor 1 (Mif1) protein positioned in the inner core of the tube of the CIS or MAC, (c) a chaperone 605 protein non-covalently associated with the Mif1 protein positioned in the inner core of the tube of the CIS or MAC, and (d) a proteinaceous cargo, or a heterologous protein or peptide, or compound, non-covalently associated or covalently associated or linked to the Mif1.