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siRNA capable of inhibiting foot and mouth disease virus replication and infection and its preparation method

A technology of foot-and-mouth disease virus and anti-foot-and-mouth disease virus, which is applied in the direction of botany equipment and methods, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve problems such as difficulties in the prevention and control of foot-and-mouth disease virus

Inactive Publication Date: 2006-04-12
FUDAN UNIV
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  • Summary
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AI Technical Summary

Problems solved by technology

This has brought great difficulties to the prevention and control of foot-and-mouth disease virus disease

Method used

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  • siRNA capable of inhibiting foot and mouth disease virus replication and infection and its preparation method
  • siRNA capable of inhibiting foot and mouth disease virus replication and infection and its preparation method

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Embodiment

[0031] Embodiment: The present invention is specifically described by taking a kind of 56POL siRNA capable of inhibiting the replication and infection of O-type FMDV in cells and animal individuals, its preparation method and application as an example.

[0032] U6 promoter of ribosomal RNA was amplified from mouse NIH / 3T3 cells by PCR.

[0033] Three gene sequences were chemically synthesized, which were: a double-stranded DNA sequence from position 1225 to position 1280 in the 3D gene of O-type FMDV polymerase, an inverted repeat sequence of the DNA sequence, and a hairpin linker DNA sequence. The three sequences are concatenated to form an inverted repeat and a siRNA template sequence connected with a hairpin linker DNA in the middle.

[0034] The siRNA template sequence was linked to the 3' end of the mouse U6 promoter to further form an siRNA expression cassette. In order to construct the recombinant adenovirus, we used the commercially available pAdTrack-CMV adenovirus s...

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PUM

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Abstract

A kind of siRNAs able to suppress the copy and infection of foot-and-mouth disease virus in cell and animal individual and its preparing process and application are disclosed. The 1D protein gene and 3D polymerase gene are chosen as the interference targets from the genome of foot-and-mouth disease virus. The human recombinant adenovirus Ad5 is designed and used as the carrier of three siRNAs. The target of 21VPI siRNA and 63VPI siRNA is said 1D gene. The target of 56POL siRNA is 3D gene.

Description

technical field [0001] The invention belongs to the technical field of foot-and-mouth disease prevention and control, and specifically relates to designing siRNA for specific sequences in the genome of foot-and-mouth disease virus (FMDV), and using adenovirus as a carrier to construct a recombinant adenovirus to express siRNA, and the siRNA can express siRNA at the cellular level and animal level (guinea pigs and pigs) ) to inhibit the infection of FMDV. Background technique [0002] Foot-and-mouth disease (FMD) is the most serious livestock infectious disease in the world today, mainly harming cloven-hoofed animals such as pigs, cattle, and sheep. For many years, foot-and-mouth disease has broken out and become popular in a large scale all over the world, causing huge economic losses to animal husbandry. According to immunogenicity, FMD virus has 7 serotypes and more than 65 subtypes, including A, O, C, SATI, SATII, SATIII and Asia I. There is no cross-immune reaction bet...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N7/01C12N15/861A61K39/235C12N15/113
Inventor 郑兆鑫严维耀陈维灶刘明秋
Owner FUDAN UNIV
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