Plant Cell Delivery Peptide Compositions for CPP Penetration
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Solution Overview
Problem
Existing cell penetrating peptides (CPPs) used in agriculture face challenges such as poor penetration efficiency and stability, limiting their effectiveness in delivering defense- and growth-promoting proteins/peptides to plant cells.
Innovation Solution
Development of peptides comprising a membrane translocation domain with specific CPP motifs and a cargo moiety, including plant bioactive components, to enhance delivery efficiency into plant cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If linear CPPs are used for plant cell delivery, then the structure is simple and easy to manufacture, but penetration efficiency is poor and stability is low
Solution Approach 1:
The CPP is divided into multiple motifs (e.g., R9 motif with 9 arginines, or combinations of shorter motifs) that can be independently optimized. Each motif contributes to membrane interaction, and their collective arrangement enhances penetration efficiency while maintaining manufacturability through modular design
Solution Approach 2:
The invention uses composite peptide structures combining multiple CPP motifs with specific cargo molecules (-defense peptides, growth promoters). This composite approach creates synergistic effects where the CPP motifs provide penetration capability while the cargo provides biological function, resolving the contradiction between simple structure and high efficiency
2Device complexity
If linear CPPs are used, then the design is simple, but proteolytic stability is poor
Solution Approach 1:
The CPP is segmented into multiple short motifs (e.g., 2-10 amino acids each) rather than one long linear sequence. This segmentation reduces the target size for proteolytic enzymes while maintaining overall penetration function, as each small motif can independently interact with membranes
Solution Approach 2:
Specific local regions (motifs) are optimized for different functions: some motifs are designed with high positive charge density for membrane binding, others with specific sequences for resistance to proteolysis. This local optimization allows the overall structure to remain relatively simple while achieving high stability
3Stability of the object's composition
If cyclic CPPs are used, then proteolytic stability improves, but chemical synthesis complexity increases
Solution Approach 1:
Rather than synthesizing one large cyclic peptide (which is complex), the invention segments the CPP into multiple small linear motifs that are easier to synthesize and then combines them. This segmentation dramatically reduces synthesis complexity while the spatial arrangement of motifs provides cyclic-like stability
Solution Approach 2:
Linker molecules serve as intermediaries connecting the CPP motifs to each other and to the cargo. These linkers facilitate modular assembly, allowing straightforward chemical synthesis of individual components that are then easily connected, avoiding the need for complex cyclic synthesis
4Productivity
If CPPs are used for delivering plant bioactive molecules, then delivery efficiency can be improved, but foliar application penetration efficiency remains poor
Solution Approach 1:
The CPP parameters are specifically optimized for plant cell membranes rather than animal cells. This includes adjusting charge density, hydrophobicity, and motif length to match plant membrane properties (e.g., presence of cutin, suberin, wax), thereby improving foliar penetration while maintaining high delivery efficiency
Solution Approach 2:
The CPP is designed as a composite structure with motifs specifically tailored for plant membrane interaction, combined with plant-specific cargo molecules (defense peptides, growth promoters). This plant-optimized composite approach resolves the contradiction by ensuring both high delivery efficiency and effective foliar penetration
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 developed peptides effectively deliver plant bioactive moieties, enhancing disease resistance and promoting plant growth, as demonstrated by increased resistance to pathogens and improved growth parameters in treated plants.
Implementation Method 1
peptides comprising a membrane translocation domain having one or more cell penetrating peptide motifs
Implementation Method 2
The first study showed that CPPs internalize into Nicotiana tabacum protoplasts, indicating that these peptides can enter plant cells by transfection
Data Source
AI summary
Described are peptides including a membrane translocation domain having one or more cell penetrating peptide motifs, and a cargo moiety linked to the membrane translocation domain, wherein the cargo moiety includes a plant bioactive moiety. The at least one cell penetrating peptide motif is from 3 to 10 amino acid residues in length and has at least three arginine and/or lysine residues; or the at least one cell penetrating peptide motif is from 3 to 10 amino acid residues in length and has at least two arginine and/or lysine residues and at least one other cell penetrating peptide motif is from 2 to 8 amino acid residues in length and has at least two hydrophobic residues. Also described are methods of delivering a cargo moiety into a plant cell comprising contacting the plant cell with the peptide as disclosed herein.