Carrier Peptide Protein Complex for Plant Cell Delivery

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Solution Overview

Problem

Current methods for introducing proteins into plant cells are complex and not extensively applicable, especially due to plant cell walls and proteolytic activity, making it difficult to achieve safe and efficient genetic modification, including transient or inheritable genome modifications.

Innovation Solution

A carrier peptide/protein complex is formed by combining a cell-penetrating sequence with a polycationic or polyanionic sequence, allowing for efficient delivery of proteins like TALEN, ZFN, or Cas9 into plant cells, which can modify genes transiently or inheritably.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If DNA transformation techniques (Agrobacterium, microparticle gun) are used to modify plants, then genetic modification can be achieved, but unexpected incorporation of foreign DNA into plant genome occurs which destructs endogenous genes

Engineering Contradiction:

Improvesafety of genetic modificationVSAvoidunexpected incorporation of foreign DNA

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts and eliminates the DNA transformation step entirely, using only protein delivery. By introducing genome-editing proteins (TALEN, ZFN, Cas9) directly into plant cells without DNA vectors, the method removes the source of unwanted foreign DNA integration while maintaining the desired genome editing function.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention uses a carrier peptide as an intermediary to deliver genome-editing proteins into plant cells. The carrier peptide forms a complex with the protein of interest, facilitates its penetration through the plant cell wall and membrane, and then releases the protein inside the cell, enabling safe protein-based genome editing without DNA transformation.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If non-transgenic direct protein delivery system is used in plants, then safety is improved, but it is difficult to achieve due to thick cell walls and proteolytic activity

Engineering Contradiction:

Improvesafety of modificationVSAvoiddifficulty of protein delivery

Core Design Contradiction:

ReliabilityVSEase of manufacture

Solution Approach 1:

The carrier peptide serves as a mediator that overcomes plant cell wall and membrane barriers. It forms a stable complex with the protein of interest, protects it from proteolytic degradation, and facilitates its penetration into the cell through the thick cell wall and plasma membrane, solving the delivery difficulty problem.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention creates a composite carrier peptide/protein complex that combines the cell-penetrating capability of the carrier peptide with the genome-editing function of the protein. This composite structure enables the protein to overcome physical barriers (cell wall, membrane) and biological defenses (proteolytic activity) that would otherwise prevent delivery.

Inventive Principle:

Principle #40Composite materials

3Productivity

If genome editing techniques (TALEN, CRISPR/Cas9, ZFN) are used to produce inheritable modification, then single treatment efficiency is improved, but it is difficult to produce transiently modified plants

Engineering Contradiction:

Improveefficiency of genome modificationVSAvoidability to produce transient modification

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

The invention makes the genome editing approach dynamic and adjustable. By delivering proteins directly rather than integrating DNA, the method allows the researcher to control whether modification is transient (protein degrades after function) or inheritable (if DNA is present during delivery). The same delivery system adapts to different outcomes based on the presence or absence of DNA templates.

Inventive Principle:

Principle #15Dynamics

Solution Approach 2:

The invention changes the key parameter from DNA-based transformation to protein-based delivery. This parameter change enables transient modification because proteins are temporary molecules that degrade after performing their function, unlike integrated DNA that is permanently inherited. The same system can achieve inheritable modification if DNA is provided as a template during the protein delivery process.

Inventive Principle:

Principle #35Parameter changes

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 method enables simple and wide-ranging protein introduction into various plant cells, facilitating safe and efficient genetic modification with controlled transient or inheritable effects.

Implementation Method 1

a carrier peptide comprising a cell-penetrating sequence and a polycationic or polyanionic sequence

Methodology Applied

Scientific EffectElectrostatic interaction: Electrostatics

Data Source

PatentUS11279941B2Method for introducing protein into plant cell

Publication Date: 2022.03.22 RIKEN CO LTD

AI summary

It is an object of the present invention to provide a method for introducing a protein into a plant, which is simple and extensively applicable to various types of plant cells and proteins. The above object is achieved by the present invention to provide a complex comprising a protein of interest to be introduced into a target plant cell and a carrier peptide, a method for introducing a protein of interest into a target plant cell using the complex, and a kit comprising a protein of interest to be introduced into a target plant cell and a carrier peptide.