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CN-121991955-A - DsRNA (double-stranded ribonucleic acid) compound auxiliary agent, and method and application thereof for preparing RNA biopesticide

CN121991955ACN 121991955 ACN121991955 ACN 121991955ACN-121991955-A

Abstract

The invention belongs to the field of biological medicine, and relates to a dsRNA (double-stranded ribonucleic acid) compound auxiliary agent, a method for preparing an RNA biological pesticide by using BioIon biological ions (BI 18) as a main material to load dsRNA, wherein the loaded dsRNA shows stronger stability in extreme environments such as RNase A, strong ultraviolet and the like, and can effectively prevent degradation of the dsRNA. Meanwhile, the dsRNA can promote the intracellular absorption of dsRNA, and has the effect of improving the expression effect of the target gene in the cotton bollworm larvae. Other aspects of the invention are designed to include these bi18@dsrna drug complexes suitable for use in the ecological arts, as well as methods of inhibiting target gene expression by administering these dsRNA drugs, for example for pest management in agriculture.

Inventors

  • GUAN RUOBING
  • WANG YINUO
  • MIAO XUEXIA
  • LI HAICHAO

Assignees

  • 上海植生优谷生物技术有限公司

Dates

Publication Date
20260508
Application Date
20260130

Claims (10)

  1. 1. Application of zinc element in preparing dsRNA compound preparation.
  2. 2. A dsRNA (double-stranded ribonucleic acid) compound auxiliary agent is characterized in that the dsRNA compound auxiliary agent is zinc salt.
  3. 3. The dsRNA compounding auxiliary agent of claim 1, wherein the zinc salt is one or more of zinc nitrate, zinc sulfate and zinc chloride.
  4. 4. The method for preparing the RNA biopesticide by using the dsRNA compounding auxiliary agent as claimed in claim 2 or 3, which is characterized by comprising the step of mixing the dsRNA compounding auxiliary agent with dsRNA under the condition of solution to form the RNA micelle biopesticide.
  5. 5. The method for preparing RNA biopesticide by using dsRNA compounding aid according to claim 4, wherein the molar ratio of the dsRNA compounding aid to the dsRNA is 1:10 -6 -1:1.
  6. 6. The method for preparing RNA biopesticide by using dsRNA compounding aid according to claim 5, wherein the content of Zn 2+ in the dsRNA compounding aid is 0.03-30% w/v.
  7. 7. The method for preparing RNA biopesticide by using dsRNA compounding aid according to claim 5, wherein the content of dsRNA in the dsRNA compounding aid is 0.5-10mg/mL.
  8. 8. The method for preparing RNA biopesticide by using dsRNA as a compounding auxiliary agent according to claim 5, wherein the dsRNA is synthesized by taking a gene with biological control effect as a template.
  9. 9. The RNA biopesticide prepared by the method according to any one of claims 5 to 8, wherein the RNA biopesticide is a cationic polyelectrolyte formed by zinc salt and dsRNA, namely dsRNA micelle.
  10. 10. Use of the RNA biopesticide of claim 9 for the preparation of a product for controlling agricultural pests.

Description

DsRNA (double-stranded ribonucleic acid) compound auxiliary agent, and method and application thereof for preparing RNA biopesticide Technical Field The invention belongs to the field of biological medicine, and relates to protection and application effectiveness improvement of dsRNA (double-stranded ribonucleic acid) serving as a core component of an RNA biological pesticide. Background The ever-deteriorating climate severely affects the adequate supply of food demand by the global population, while the worsening climate results in irregular outbreaks of some pests, estimated to be up to 40% of the global crop loss per year due to pests, economic losses up to $ 3000 billion, and new or repeated outbreaks of pests in crop areas accompanying food shortages and high density rapid population growth. Based on plant genetic resistance and/or transgene, chemical/biological agents, and natural enemy applications, although crop yield is greatly improved, problems such as soil degradation, resistance, residue, and ecological chain breakage are also solved, which poses a threat to personal, national, and even global, grain safety, ecological safety, and the like. However, even so, the safe and adequate supply of crops is not effectively protected. The RNA biological pesticide brings about another revolution in pesticide history, and is a novel biological pesticide developed based on RNA interference technology, and the core component of the novel biological pesticide is double-stranded RNA. After entering an organism, the exogenous double-stranded RNA is digested into small interfering RNA, the small interfering RNA is combined with an RNA-induced silencing complex composed of multiple molecules, and an antisense strand of the small interfering RNA finds an information RNA sequence complementary with a base of the small interfering RNA and cuts and degrades the information RNA sequence, so that normal translation of target proteins is affected, and further the function of the organism is disturbed, so that the growth and development of the organism are blocked, and even the organism dies, thereby achieving the aim of preventing and controlling diseases and insect pests. In addition, by taking computational biology, synthesis biology and materiality as assistance, the RNA pesticide takes a brand new action mechanism, takes prevention and control precision, low cost, simple operation and ecological safety as a core, basically meets most of the expectations of people on green pesticide core elements, and provides a brand new solution for high-quality sustainable agricultural pest control. The application mode of the RNA interference technology in agricultural pest control mainly comprises plant source RNA pesticides and spray RNA pesticides. The Plant source RNA pesticide, also called Plant-incorporated Protectant, PIP, is dsRNA for expressing Plant pest target gene in Plant, and the spray RNA pesticide, called non-PIP, is based on exogenous synthesized dsRNA effective to some Plant pest, and is applied by spraying directly after being made into preparation, and publication No. CN121065189A discloses dsRNA for preventing and controlling leaf spot miner and its pesticide compound. Despite its widespread use and intensive research into RNAi mechanisms, differences in the nature of double-stranded RNA (dsRNA) uptake, systemic transmission, sensitivity of insects to dsRNA, have resulted in the difficulty of effective application of naked dsRNA to a part of the world's major agricultural pests such as cotton bollworms. Thus, there is a need for new dsRNA stabilizers and delivery aids to facilitate field application of RNAi technology. Disclosure of Invention In order to solve the technical problems of stability and use effect of dsRNA, the invention provides a dsRNA compound auxiliary agent, and a method for preparing RNA biopesticide by using the same. The technical scheme of the invention is realized as follows: In one aspect, the application provides a dsRNA formulation aid, wherein the dsRNA formulation aid bioion18 (BioIon, BI 18) is a zinc salt, including but not limited to zinc nitrate (Zn (NO 3)2), zinc sulfate (ZnSO 4), zinc chloride (ZnCl 2), and the like. In a second aspect, the invention provides a method for preparing RNA biopesticide by using the dsRNA compounding auxiliary agent, which comprises the steps of mixing the dsRNA compounding auxiliary agent with dsRNA under the condition of solution to obtain the RNA biopesticide. BI18 is capable of payload dsRNA, increasing the stability and drug loading of the BI18 and dsRNA complex by intercalation, exoglobus complexation, endoglobus complexation of BI18, buffering of pH by Tris-HCl. BI18 can further compress dsRNA to obtain BI18@dsRNA micelle with a certain rigid structure. The compound can protect dsRNA from enzymolysis of RNase A in the range of a molar ratio of 1:10 -6–1:10-3 and can resist extreme ultraviolet in the range of a molar ratio of 1:10 -3–1:10-1.