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Application of siRNA sequences and target thereof in improving PEDV titer

A technology of sequence and toxicity, which is applied in the application field of siRNA sequence and its target in improving the toxicity of PEDV, which can solve the problems of complicated downstream signaling pathways and complex LncRNA structure.

Pending Publication Date: 2020-10-23
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
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Problems solved by technology

[0004] However, due to the complex structure of LncRNA and the cumbersome downstream signaling pathways, the regulation of LncRNA will produce a series of complex chain reactions, which makes the existing technology unrealized. By regulating the lncRNA of Vero cells to increase the toxicity of PEDV in Vero cells, This concept is still blank in this field

Method used

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  • Application of siRNA sequences and target thereof in improving PEDV titer
  • Application of siRNA sequences and target thereof in improving PEDV titer
  • Application of siRNA sequences and target thereof in improving PEDV titer

Examples

Experimental program
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Embodiment 1

[0058] 1. Expression of LNC_002015 in Vero-E6 cells at different infection time periods

[0059] Taking Vero-E6 cells infected with PEDV with MOI=0.01 as the model, the cells in the time period with the highest intracellular virus titer were selected as the experimental group, and the uninfected cells in the same time period were used as the blank control, and the second-generation full-transcriptome high-throughput Sequencing, after analyzing and processing the sequencing data, the Vero-E6 cells were screened out to significantly up-regulate the expression of LNC_002015, and the expression in different time periods was detected (the primers for detecting LNC_002015 are shown in Table 1).

[0060] Table 1. Detection Primers and Probes

[0061]

[0062] The experimental group was Vero-E6 cells inoculated with CV777 at a multiplicity of infection MOI=0.01, and the blank control group was the control group of Vero-E6 cells not inoculated with CV777. After inoculation, 12, 24, ...

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Abstract

The invention discloses two siRNA sequences and target gene sequences thereof. Two target spots of si-LNC_002015 gene in Vero-E6 culture cells are interfered through siRNA, so that the PEDV RNA copy number and the protein expression quantity in the Vero-E6 culture cells are obviously increased compared with those of a control group, and the virus titer is increased.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to the application of an siRNA sequence and its target in increasing the virulence of PEDV. Background technique [0002] Porcine Epidemic Diarrhea Virus (PEDV) is a virus that can infect pigs of all ages in the αcoronavirus genus, and can cause acute, highly contagious and destructive intestinal diseases in pigs, especially 1- The morbidity rate of 2-week-old piglets is as high as 100%, and the mortality rate can be as high as 80%-100%, which has caused serious economic losses to the global pig industry, including my country, pushed up meat prices, and affected people's living standards. The use of an effective PEDV vaccine is of great significance for blocking the spread of the virus. The Vero cell line is an ideal cell line for large-scale production of PEDV commercial vaccines. The titer of PEDV replicated in Vero cells will directly affect the cost of vacci...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N5/10C12N7/00
CPCC12N15/113C12N7/00C12N5/0686C12N2310/14C12N2510/00C12N2770/20051Y02A50/30
Inventor 兰喜李学瑞王志林杨彬吴锦艳曹小安尚佑军刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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