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Method for interfering gene expression of insect through infiltrating dsRNA through insect body walls

A gene expression and insect technology, applied in the field of nanomaterials and gene interference, can solve problems such as difficult to eliminate insect errors, insect tissue injury, body fluid loss, etc., to achieve the effect of reducing the loss of dsRNA molecules, no operation trauma, and promoting adhesion

Pending Publication Date: 2018-06-15
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] But even so, there is still the problem that the microinjection method can easily cause insect internal tissue injury, body fluid loss or even death, and the feeding method is difficult to rule out the error caused by malnutrition of insects

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] This embodiment is used to illustrate the preparation method of the complex medicinal liquid of the present invention, specifically as follows:

[0041] 1. Raw materials:

[0042] (1) Nanocarrier: In this embodiment, the fluorescent nanocarrier molecule disclosed in the patent No. 201310119837.3 is taken as an example. The fluorescent nucleus inside the carrier has a high fluorescence quantum yield, good light, heat and chemical stability, and is conducive to in vivo Tracking, the formation of a good cavity structure between each side chain and a large number of specific functional groups are conducive to the binding and assembly of dsRNA molecules, and form a stable dsRNA molecule / carrier complex. The whole complex has good insect cell affinity It is beneficial to the rapid entry of dsRNA molecules into cells to target target genes.

[0043] (2) dsRNA: dsRNA that interferes with genes synthesized in the fat body.

[0044] According to the standard method for in vitro...

Embodiment 2

[0053] This example is used to illustrate how to use the complex drug solution described in Example 1 to deliver dsRNA into small piercing-sucking mouthparts insect soybean aphid, interfere with the expression of key genes and control soybean aphid oviposition.

[0054] In this example, the soybean aphid is used as a representative tested insect. Its abdomen is full of body fluid, and microinjection of dsRNA molecules causes the internal tissue injury of soybean aphids, and most of the body fluids are lost and dies; herbivorous soybean aphids lack nutritious and comprehensive artificial diets, and it is difficult to rule out that the tested aphids are caused by malnutrition by feeding soybean aphids with dsRNA molecules. The error; its volume is small, the body length is about 2 μm, and it is easy to be drowned by the large-volume drug solution with insufficient atomization during the application process. In this example, the soybean aphid, which is relatively difficult to ope...

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Abstract

The invention relates to the fields of nano-materials and gene interference, and concretely discloses a method for interfering the gene expression of an insect by infiltrating dsRNA through insect body walls. The adhesion, distribution and infiltration of exogenous dsRNA molecules to an insect body surface and entrance of the molecules to various internal organ tissues are realized by means of theeffect of a surfactant by complexing a nano-vector with a dsRNA functional macromolecule in order to efficiently interfere the gene expression. The efficient and noninvasive introduction of the exogenous dsRNA molecules to insect body cells by beans of the dual synergistic effects of the nano-vector and the surfactant in order to finally promote the correct targeting of the dsRNA molecules and improve the RNA interference rate. The method has the advantages of simplicity, high efficiency and small error, and is of great practical significance to the scientific research of the gene interference of the insect and the application of efficient and safe targeting RNA pesticide preparations.

Description

technical field [0001] The invention relates to the fields of nanomaterials and gene interference, in particular to a method for interfering gene expression of insects by penetrating dsRNA through insect body walls. Background technique [0002] RNA interference (RNA interference, RNA interference) refers to the phenomenon of efficient and specific degradation of homologous mRNA induced by double-stranded RNA (double-stranded RNA, dsRNA molecule), which is highly conserved during evolution. These specially designed exogenous dsRNA molecules can specifically, efficiently and permanently reduce the expression of specific genes, which is of great significance for studying the functions of unknown genes, regulating the development and behavior of insects, and protecting plants from viruses. As an important method, RNA interference has been widely used in the identification of various gene functions and the regulation of functional gene expression. At the same time, due to the h...

Claims

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Application Information

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IPC IPC(8): C12N15/87A01K67/033
CPCA01K67/033A01K2227/706C12N15/87
Inventor 沈杰郑洋王丹刘小侠黎剑浩尹梅贞
Owner CHINA AGRI UNIV
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