Peptide-Conjugated PPMOs for Targeted RNA Silencing in Plant Pathogens
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Solution Overview
Problem
Current methods for targeting plant pathogens and insect pests, particularly bacterial infections in crops, face challenges such as antibiotic resistance, cost, and safety concerns for beneficial microbes, necessitating the development of non-antibiotic compositions and methodologies for effective and specific treatments.
Innovation Solution
The use of peptide-conjugated phosphorodiamidate morpholino oligomers (PPMOs) that target specific bacterial genes, allowing for RNA silencing in bacteria within plants and insects, thereby controlling bacterial infections through ingestion of oligonucleotides by insects or direct application to plants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If antibiotics are used to treat bacterial plant pathogens, then treatment effectiveness is improved, but antibiotic resistance develops and beneficial microbes are harmed
Solution Approach 1:
The invention changes the fundamental parameter of the treatment mechanism from antibiotic-based bacterial killing to RNA interference-based gene silencing. PPMOs target specific bacterial mRNA sequences to inhibit protein translation, thereby changing the mode of action from broad-spectrum antibacterial to highly specific gene suppression, avoiding antibiotic resistance while preserving beneficial microbes
Solution Approach 2:
The invention applies local quality by designing PPMOs with sequence-specific complementarity to target bacterial mRNA. Each PPMO is tailored to bind only to its specific target sequence, enabling localized and specific gene silencing in the pathogen without affecting other bacteria or beneficial microbes in the ecosystem
2Reliability
If conventional chemical treatments are used to control plant pathogens and insect pests, then disease control is improved, but safety concerns arise for beneficial microbes and environmental impact increases
Solution Approach 1:
The invention uses sequence-specific PPMO designs that target only the intended pathogen or pest through complementary base pairing with their unique mRNA sequences. This local specificity ensures that beneficial microbes are not affected, as the PPMOs do not bind to non-target organisms
Solution Approach 2:
The invention introduces PPMOs as intermediary molecules that mediate gene silencing in target organisms. These oligonucleotides act as specific mediators that can be delivered to plants, insects, or mites, where they selectively silence genes in the target pathogen or pest without broadly affecting the ecosystem
3Productivity
If peptide conjugation is added to PMOs to enhance cellular uptake, then delivery efficiency is improved, but molecular complexity increases
Solution Approach 1:
The invention merges two functional components: the PMO molecule for gene silencing and the cell-penetrating peptide for enhanced delivery. This combination creates a conjugate where the peptide portion facilitates cellular uptake while the PMO portion maintains its gene-silencing function, achieving synergistic improvement in delivery efficiency
Solution Approach 2:
The cell-penetrating peptide component provides multi-functionality by enabling delivery across different cell types and biological barriers. This universal delivery mechanism allows the PPMO conjugate to effectively reach intracellular targets in diverse organisms including plants, insects, and mites
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
PPMOs effectively induce RNA silencing in bacterial pathogens, reducing their populations and causing detrimental effects in insects, thereby controlling bacterial infections in plants and insects, offering a specific and safe alternative to traditional antibiotics.
Implementation Method 1
ASOs act through several different mechanisms and have a wide variety of structures and names. The present disclosure focuses on PPMOs in particular... PMOs form stable base pairs with complementary nucleic acid sequences
Implementation Method 2
The PMO backbone is made of morpholino rings with phosphorodiamidate linkage, which protects them from nuclease degradation while still maintaining the complementary base pairing
Implementation Method 3
Covalent conjugation of the PMO with membrane-penetrating peptides—resulting in protein-conjugated PMOs (PPMOs)—can enhance their cellular uptake by mammalian cells as well as bacteria
Implementation Method 4
Antisense phosphorodiamidate morpholino oligomers (PMOs) downregulate target gene expression by interfering with the binding of ribosome to mRNA and thereby inhibiting protein translation
Data Source
AI summary
The current disclosure provides for peptide-conjugated phosphorodiamidate morpholino oligomers (PPMOs) antisense oligonucleotides that target plant pathogens and insect pests by targeting the microbes within the insect pest. Methods of delivering PPMOs to bacteria in plants, in insect carriers, and to the insects via plant feeding are also provided.