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RNA disturbance target point for inhibiting MDV proliferating and application thereof

An RNA interference and target technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, sugar derivatives, etc., can solve the problem of unclear exact mechanism, and achieve the effect of convenient enzyme digestion and identification

Inactive Publication Date: 2011-08-10
上海市农业科学院畜牧兽医研究所
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the exact mechanism of RNAi effect is not clear. It is generally believed that dsRNA is cut into 19-23bp small interference RNA (small interference RNA, siRNA, siRNA combines with some enzymes in the body (including endonuclease, exonuclease, helicase) to form an RNA-induced silencing complex RISC (RNA induced silencing complex, RISC). RISC then specifically binds to the homologous region of mRNA, Enzymatically degrades mRNA and silences related genes (gene silence)

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  • RNA disturbance target point for inhibiting MDV proliferating and application thereof
  • RNA disturbance target point for inhibiting MDV proliferating and application thereof
  • RNA disturbance target point for inhibiting MDV proliferating and application thereof

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Embodiment Construction

[0019] In the following, the present invention will be further described in detail in combination with the experiments of different siRNAs inhibiting the infection of CEF cells by MD virus.

[0020] 1. Experimental method

[0021] 1.1 siRNA DNA template design: According to the experimental experience, using biological software, according to the gene sequence of MDV (virus strain Md5) published by GENEBANK, designed and prepared 6 siRNAs, the targeted gene is the viral cortex protein expressed in the MDV virus UL49 gene sequence of Vp22. Finally, the BLAST software provided by NCBIGenBank (written by the National Library of Medicine and the National Institutes of Health) was used to perform homology analysis on the selected target sequences to exclude the possibility of non-specific inhibition of chicken and other viral gene fragments by siRNA. pSilencer TM 2.1-U6 neo vector has U6 promoter, which can transcribe siRNA. Designed to be cloned into pSilencer TM 2.1-U6 neo v...

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Abstract

The present invention belongs to the field of molecular biology and preventive veterinary medicine, and discloses one group of RNA interfere target points on MDV and MDV treating preparations with siRNA or antisense RNA obtained based on these target sequences. Through initial screening based on the available results and transfecting cell with siRNA expression vector, six RNA interfere target points on MDV genome is screened out, with the No. 4 target point having MDV proliferation inhibiting rate as high as 97 %. The present invention relates to the acting site for gene therapy, and may be applied in screening new medicines for preventing and treating MDV infection.

Description

technical field [0001] The invention belongs to the fields of molecular biology and preventive veterinary medicine, and particularly relates to a set of targets for RNA interference against MDV, and obtaining siRNA or antisense RNA according to these target sequences for preparing preparations for treating MDV. Background technique [0002] Marek's Disease (MD) is a highly contagious lymphoproliferative disease of chickens caused by Marek's Disease Virus (MDV). MD is widely prevalent all over the world, and it is one of the most serious infectious diseases that harm the poultry industry at present, causing huge economic losses to the poultry industry. Judging from the history of MD development and the current situation of prevention and control, it is not enough to control MD only by vaccines. Many factors, such as early infection and virus virulence evolution, can cause immune failure. MD experts and researchers from all over the world are constantly researching and discus...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C07H21/02A61K48/00A61P31/14
Inventor 缪德年王秀花樊生超姚惠娟
Owner 上海市农业科学院畜牧兽医研究所