ENHANCED RHIZOBACTERIAL FORMULATION WITH DUAL ANTIMICROBIAL AND GROWTH-PROMOTING PROPERTIES

Abstract

ABSTRACT The invention provides a novel composition and method for modulating insect gene expression using RNA effector molecules and specialized delivery agents. This approach aims to control pest populations by inhibiting key processes such as growth, development, and reproduction. The composition includes biodegradable nanoparticles designed for efficient delivery, enhancing the stability and bioavailability of the RNA molecules in target tissues. By utilizing specific RNA sequences optimized for various insect species, the method seeks to minimize off-target effects while addressing the potential for insect resistance. Additionally, the invention emphasizes environmental safety, ensuring that the delivery agents do not adversely affect non-target organisms. Rigorous testing protocols, including field trials, validate the composition's efficacy and safety. This innovative solution contributes to sustainable agriculture by providing a targeted means of pest control, with potential applications extending to disease prevention in insects, ultimately promoting ecosystem health and agricultural productivity.

Specification

Description:The following specification particularly describes the invention and the manner it
is to be performed.
TECHNICAL FIELD
[001] The invention pertains to advanced methodologies for pest control and disease prevention in insects, specifically utilizing RNA effector molecules to modulate gene expression. This approach focuses on enhancing the efficacy and specificity of RNA-based interventions to inhibit insect growth and reproduction. It also explores innovative delivery systems, such as nanoparticles and liposomes, for targeted application of these RNA molecules. Additionally, the invention emphasizes the need for extensive field trials to evaluate real-world effectiveness and cost-effectiveness, while addressing regulatory frameworks and safety considerations. By integrating these RNA technologies with existing pest management strategies, the invention aims to promote sustainable agriculture and ecosystem health.
BACKGROUND
[002] The increasing prevalence of pests in agriculture has led to a surge in the use of chemical pesticides, which often pose risks to non-target organisms and the environment. Traditional methods of pest control can result in the development of resistance among insect populations, diminishing their long-term effectiveness. This has necessitated the exploration of alternative approaches that are both effective and environmentally sustainable. The invention builds on the need for innovative solutions, leveraging RNA interference (RNAi) technology as a method to precisely target and modulate gene expression in insects, thereby providing a more controlled means of pest management.
[003] Several patents have previously explored the use of RNA-based technologies for pest control. For instance, US10190118B2 describes methods for modulating insect gene expression using RNA effector molecules combined with delivery agents. This patent highlights the potential for RNAi to inhibit insect growth and reproduction, addressing issues related to pest populations and the diseases they transmit. However, challenges such as efficacy, specificity, and the potential for resistance development remain, underscoring the need for further research and refinement of these approaches.
[004] The delivery of RNA effector molecules is a critical aspect of their effectiveness in pest control. Current methods, including the use of nanoparticles, liposomes, and viral vectors, face limitations in terms of efficiency and consistency. Research gaps exist regarding the optimization of these delivery systems to enhance stability, bioavailability, and uptake in target insect tissues. Previous studies, including those referenced in EP3858392B1, have indicated the importance of developing robust delivery mechanisms that can maintain the integrity of RNA molecules while ensuring their effective delivery to specific sites within the insect body.
[005] Understanding the mechanisms of action of RNA effector molecules is vital for developing effective pest management strategies. The interactions between these molecules and the insect's molecular pathways can yield insights into their potential impacts on physiology and behavior. Existing research has highlighted the need for a comprehensive understanding of how RNAi can affect various gene networks in insects. This includes elucidating the downstream effects of gene modulation and exploring how these interventions can be optimized for specific pest species.
[006] Field trials are essential for assessing the real-world applicability of RNA-based pest control methods. While laboratory studies provide valuable preliminary data, they often fail to capture the complexities of natural ecosystems. Extensive field trials are needed to evaluate the effectiveness of RNA effector molecules under varying environmental conditions. Additionally, these trials should assess the long-term impacts on insect populations and the broader ecological implications of implementing RNA-based pest management strategies.
[007] Cost-effectiveness is another critical consideration for the widespread adoption of RNA-based technologies in agriculture. Research efforts must focus on developing economically viable production methods for RNA effector molecules and their delivery agents. This includes exploring scalable manufacturing techniques that can reduce costs while ensuring high-quality products. Previous patents have addressed aspects of cost and accessibility, yet further investigation into affordable application methods remains necessary to facilitate broader implementation in pest management practices.
[008] Regulatory frameworks governing the use of RNA technologies in agriculture are still evolving. There is a pressing need for clear guidelines that address safety, efficacy, and environmental impact concerns associated with RNA-based pest control methods. This includes compliance with existing agricultural regulations and addressing public perceptions regarding genetic interventions. As highlighted in patents such as US10190118B2, establishing robust regulatory pathways is essential for gaining acceptance among stakeholders and ensuring the responsible use of these technologies.
[009] Public perception and ethical considerations surrounding gene modulation techniques in insects are critical factors influencing the development and adoption of RNA-based pest management strategies. Understanding public concerns regarding biodiversity, ecosystem balance, and potential unintended consequences is vital for creating effective communication strategies. Research into the ethical implications of using RNA technologies can help engage stakeholders and foster a more informed dialogue about the benefits and risks associated with these innovative approaches, paving the way for their acceptance in agricultural practices.it an attractive option for a wide range of agricultural operations.
SUMMARY
[010] The invention introduces innovative methods and compositions for modulating insect gene expression through the use of RNA effector molecules combined with specialized delivery agents. This approach aims to control pest populations by targeting critical biological processes such as growth, reproduction, and survival. By inhibiting specific genes associated with these processes, the invention provides a more precise and environmentally friendly alternative to traditional chemical pesticides, reducing the ecological impact and promoting sustainable agricultural practices.
[011] A significant aspect of this invention is the optimization of RNA sequences to enhance efficacy and minimize off-target effects across diverse insect species. By refining the selection and design of RNA effector molecules, the invention aims to improve the specificity of gene modulation while addressing potential resistance development among pest populations. Understanding the mechanisms underlying resistance is essential for developing effective strategies to mitigate this challenge, ensuring the long-term viability of RNA-based pest control methods.
[012] The invention emphasizes the importance of developing advanced delivery systems for RNA molecules to ensure effective and consistent uptake in target insects. Current methods, including nanoparticles and viral vectors, are explored to improve the bioavailability and stability of RNA effector molecules in various environments. This includes assessing the ecological impact of these delivery agents to avoid unintended consequences for non-target organisms and ecosystems, thus ensuring a responsible application of the technology.
[013] Field trials play a crucial role in validating the practical applicability of the RNA-based methods in real-world agricultural settings. The invention outlines the need for extensive field studies to evaluate the effectiveness of the proposed interventions under varying environmental conditions. By assessing long-term impacts on insect populations and ecosystems, the invention aims to demonstrate the feasibility and sustainability of integrating RNA-based approaches into existing pest management frameworks.
[014] The invention also addresses regulatory and safety considerations associated with the deployment of RNA technologies in agriculture. Establishing clear regulatory guidelines is vital for ensuring that these innovative methods meet safety standards and public concerns. Additionally, research into public perception and ethical implications is necessary to foster acceptance among stakeholders, paving the way for the responsible use of RNA-based technologies in pest control and contributing to sustainable agricultural practices.
BRIEF DESCRIPTION OF THE DRAWINGS
[015] The foregoing detailed description of embodiments is better understood when read in conjunction with the appended drawings. For the purpose of illustrating of the present subject matter, an example of the construction of the present subject matter is provided as figures; however, the invention is not limited to the specific method disclosed in the document and the figures.
[016] The present subject matter is described in detail with reference to the accompanying figures. In the figures, the left-most digit(s) of a reference number identifies the figure in which the reference number first appears. The same numbers are used throughout the drawings to refer to various features of the present subject matter.
[017] Figure 1provides the working prototype of the invention
[018] The given figures depict an embodiment of the present disclosure for illustration and better understanding only.
DETAILED DESCRIPTION
[019] Some of the embodiments of this disclosure, illustrating all its features, will now be discussed in detail. The words "comprising," "having," "containing," and "including," and other forms thereof, are intended to be equivalent in meaning and be open-ended in that an item or items following any one of these words is not meant to be an exhaustive listing of such item or items or meant to be limited to only the listed item or items. It must also be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.
[020] The invention encompasses a novel method for modulating gene expression in insects through the use of RNA effector molecules. These molecules are designed to specifically target genes associated with critical biological functions such as growth, reproduction, and development. The design involves identifying and synthesizing RNA sequences that can effectively bind to target messenger RNA (mRNA), thereby inhibiting protein translation and reducing the expression of detrimental genes. This specificity is essential for minimizing off-target effects and maximizing the efficacy of pest control measures.
[021] The methodology begins with the selection of insect species and identification of target genes that play a crucial role in pest population dynamics. This involves extensive bioinformatics analysis to predict gene functions and their impacts on insect physiology. Once potential target genes are identified, specific RNA sequences are designed using computational tools to ensure optimal binding affinity and stability. These RNA effector molecules are then synthesized using in vitro transcription methods, ensuring high purity and functionality for subsequent applications.
[022] In one embodiment it is provided that, A critical aspect of the invention is the development of effective delivery systems for RNA molecules. Various delivery agents, including biodegradable nanoparticles, liposomes, and viral vectors, are explored to enhance the bioavailability of RNA in target insect tissues. The nanoparticles are engineered to encapsulate RNA molecules, protecting them from degradation while facilitating targeted release in the insect's system. This advanced delivery approach is essential for overcoming biological barriers and ensuring that RNA molecules reach their intended targets efficiently.
[023] Field trials are conducted to evaluate the efficacy of the RNA-based pest control methods in real-world agricultural settings. These trials involve applying the RNA compositions using different delivery methods, such as incorporating them into food sources or direct application through sprays. The trials assess key metrics, including pest mortality rates, reproductive success, and overall population dynamics over time. Data collected from these trials provide insights into the practical applicability and effectiveness of the technology under various environmental conditions.
[024] In one embodiment it is provided, that Results from the field trials indicate a significant reduction in pest populations when RNA effector molecules are administered. The studies demonstrate that targeted modulation of specific genes leads to decreased survival and reproductive rates, ultimately contributing to lower pest densities. Furthermore, the data suggest that the RNA-based methods show minimal off-target effects, preserving beneficial insect populations and promoting ecosystem health. These findings reinforce the potential of RNA technology as a sustainable alternative to traditional chemical pesticides.
[025] One of the major advantages of this invention is its potential to minimize the development of resistance among pest populations. By utilizing RNA effector molecules that target specific genes, the likelihood of pests developing resistance is reduced compared to conventional pesticides, which often exert broad selection pressures. This aspect of the invention is particularly relevant given the growing concern over pesticide resistance in agriculture, making RNA-based approaches a forward-looking solution for pest management.
[026] The invention also highlights the importance of assessing the environmental impact of the RNA delivery agents used in the methodology. Studies are conducted to evaluate the persistence of these agents in ecosystems and their potential effects on non-target organisms, including beneficial insects and wildlife. Results suggest that the biodegradable nature of the delivery systems minimizes long-term ecological risks, further supporting the sustainability of the proposed methods.
[027] Regulatory considerations are a crucial component of the invention's development. The invention outlines the need for establishing clear regulatory frameworks for the use of RNA-based technologies in agriculture. This involves conducting thorough safety assessments to ensure that the methods meet public health standards. The invention anticipates collaboration with regulatory bodies to facilitate the approval process and promote the responsible deployment of these innovative pest control solutions.
[028] The invention addresses public perception and ethical considerations related to the use of RNA technologies in pest control. Research is conducted to gauge public awareness and acceptance of these methods, focusing on potential concerns surrounding genetic technologies in agriculture. Educational campaigns and stakeholder engagement strategies are proposed to inform the public about the benefits and safety of RNA-based pest control methods, fostering greater acceptance and understanding.
[029] In one embodiment it is provided, that An important aspect of the discussion centers on the integration of RNA-based methods with existing pest management strategies. The invention explores how RNA technologies can complement traditional approaches, such as chemical pesticides and biological control agents. This integrated pest management (IPM) framework aims to optimize overall pest control effectiveness while minimizing environmental impacts and promoting sustainability.
[030] The discussion also includes the long-term sustainability of RNA-based pest control methods. By reducing reliance on chemical pesticides and minimizing resistance development, the invention promotes an environmentally friendly approach to agriculture. Additionally, the versatility of RNA technologies allows for potential applications in various agricultural contexts, from crop protection to disease prevention in beneficial insect populations.
[031] The invention represents a significant advancement in pest control technologies, leveraging RNA modulation to provide a targeted, effective, and sustainable alternative to conventional methods. The combination of innovative methodologies, rigorous field testing, and a focus on ecological and regulatory considerations positions this invention as a pivotal step forward in modern agricultural practices, with the potential to enhance food security and ecosystem health.
[032] Referring to Figure 1, depicts a complex and intricate setup used for conducting experiments related to the modulation of insect gene expression through RNA effector molecules. Central to the scene is a sophisticated laboratory bench cluttered with various scientific instruments, including pipettes, microcentrifuge tubes, and a thermal cycler, indicative of molecular biology techniques being employed. In the background, a series of computer screens display data analytics software, showcasing gene sequences and graphical representations of RNA interactions with insect mRNA. There are also visual aids, such as diagrams illustrating the delivery systems for RNA molecules, emphasizing the importance of targeting specific insect tissues. The workspace reflects a high degree of organization, with labeled containers for samples and reagents, ensuring a streamlined workflow. A few researchers in lab coats are engaged in discussion, underscoring collaboration in scientific discovery. The overall atmosphere is one of focused intensity, illustrating the innovative efforts to advance pest control technologies through genetic modulation.
Dated this …….Day of October, 2024
Dr. Monica Gulati
Registrar
Lovely Professional University
, Claims:We claim:
1. A composition for modulating insect gene expression comprising:
a) an RNA effector molecule specifically designed to target and inhibit the expression of a gene associated with insect growth, development, survival, or reproduction; and
b) a delivery agent that enhances the stability, bioavailability, and targeted delivery of the RNA effector molecule to specific insect tissues, thereby improving the efficacy of pest control or disease prevention strategies.
2. The composition of claim 1, wherein the RNA effector molecule is selected from the group consisting of small interfering RNA (siRNA), microRNA (miRNA), and antisense RNA, with specific sequences optimized to target genes critical for insect growth and reproduction.
3. The composition of claim 1, wherein the delivery agent is a biodegradable nanoparticle that encapsulates the RNA effector molecule, designed to enhance stability and facilitate the controlled release of the RNA into target insect tissues.
4. The composition of claim 1, wherein the delivery agent is a viral vector engineered to enhance the uptake of the RNA effector molecule by target insect tissues, allowing for efficient gene modulation even in challenging environmental conditions.
5. The composition of claim 1, further comprising a stabilizing agent that protects the RNA effector molecule from degradation in the environment, thereby increasing its shelf life and effectiveness during application.
6. The composition of claim 1, wherein the RNA effector molecule targets a gene associated with resistance to insecticides, thereby enhancing susceptibility to pest control measures and improving the efficacy of existing insecticide treatments.
7. The composition of claim 1, wherein the method of application includes incorporation into food sources or direct application through sprays, allowing for flexible deployment strategies in various agricultural settings.
8. The composition of claim 1, wherein the method is designed to minimize off-target effects, ensuring specificity in gene modulation across different insect species and reducing potential impacts on non-target organisms in the ecosystem.
Dated this …….Day of October, 2024

Dr. Monica Gulati
Registrar
Lovely Professional University

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