Protein Transduction Delivery Molecule for Safe Nuclear Cargo Uptake

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Solution Overview

Problem

Current methods for protein transduction into cells are inefficient, as they rely on DNA constructs that can cause genetic modifications and insertional mutagenic lesions, and existing protein delivery systems struggle to achieve physiological or metabolic effects due to endosomal retention and low protein escape from macropinocytotic vesicles.

Innovation Solution

A delivery molecule comprising a cargo, a glycosaminoglycan (GAG) binding element, and a protein transduction domain (PTD) that facilitates the uptake and nuclear delivery of proteins into cells, enhancing transduction efficiency by binding to cell surface GAGs and using macropinocytosis for cellular entry.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If DNA constructs are used for protein transduction, then gene expression can be achieved, but insertional mutagenic lesions and genetic modifications occur

Engineering Contradiction:

Improvesafety of transduction methodVSAvoidinsertional mutagenic lesions

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts and eliminates the DNA construct component from the transduction system, using only protein-based delivery molecules (peptides or proteins) to achieve transduction without genetic material, thereby removing the source of insertional mutagenesis while maintaining delivery functionality

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention introduces protein transduction domains (PTDs) and GAG-binding elements as intermediary molecules that mediate cellular uptake and nuclear delivery of cargo proteins without requiring DNA constructs, serving as a safe bridge between extracellular delivery and intracellular target engagement

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If conventional protein delivery systems are used, then protein can be delivered into cells, but endosomal retention prevents physiological or metabolic effects

Engineering Contradiction:

Improvefunctional efficacy of delivered proteinVSAvoidprotein escape from macropinocytotic vesicles

Core Design Contradiction:

ReliabilityVSQuantity of substance

Solution Approach 1:

The invention introduces nuclear localization signals (NLS) as intermediary elements that guide cargo proteins from macropinocytotic vesicles directly to the nucleus, bypassing endosomal degradation pathways and ensuring functional delivery of the protein cargo to its target compartment

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention changes the destination parameter of protein delivery from cytosolic/macropinocytotic vesicle retention to nuclear localization by incorporating NLS sequences, thereby altering the subcellular distribution parameter to achieve physiological or metabolic effects

Inventive Principle:

Principle #35Parameter changes

3Reliability

If protein transduction is performed without DNA constructs, then genetic safety is improved, but transduction efficiency is low

Engineering Contradiction:

Improvesafety of transduction methodVSAvoidprotein delivery efficiency

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The invention creates composite delivery molecules by fusing multiple functional domains (GAG-binding elements, PTDs, NLS sequences, and cargo proteins) into single polypeptide constructs, combining the advantages of each component to achieve both high efficiency and safety

Inventive Principle:

Principle #40Composite materials

Solution Approach 2:

The invention merges multiple previously separate delivery mechanisms (GAG-binding for cell surface attachment, PTD for membrane transduction, NLS for nuclear targeting) into unified fusion proteins, synergistically enhancing overall delivery efficiency while maintaining the DNA-construct-free safety profile

Inventive Principle:

Principle #5Merging (Combining)

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 achieves a >100-fold improvement in protein delivery efficiency compared to existing systems, allowing for physiological or metabolic effects and bypassing the need for DNA constructs, with the GAG binding element and PTD synergizing to facilitate high-level protein uptake and nuclear delivery.

Implementation Method 1

a glycosaminoglycan (GAG) binding element, which is capable of binding to GAG on the surface of the cell

Methodology Applied

Scientific EffectGlycosaminoglycan binding: Adsorption

Implementation Method 2

using macropinocytosis for cellular entry

Methodology Applied

Scientific EffectMacropinocytosis:

Data Source

PatentUS11672870B2Transduction

Publication Date: 2023.06.13 UNIVERSITY OF NOTTINGHAM

AI summary

This invention relates to transduction of cargo molecules into living cells, such as protein transduction, in particular a delivery molecule for transduction of a cargo into a cell comprising: a cargo-binding molecule and/or a cargo; a glycosaminoglycan (GAG) binding element, which is capable of binding to GAG on the surface of the cell; and a protein transduction domain. Methods of transduction, methods of producing or modifying cargo for transduction, delivery molecules for transduction and methods of treatment using transduction, or using transduced cells are also provided.