Method for preparing foot-and-mouth disease virus-resistant RNAi transgenic livestock

Abstract

The invention discloses a method for preparing foot-and-mouth disease virus-resistant (FMDV) transgenic livestock by expressing FMDV-resistant ribonucleic acid interference (RNAi), which comprises the following steps of: linearizing an FMDV-resistant RNAi recombinant Lenti-virus vector which carries a short hairpin RNA (shRNA) base sequence and a green fluorescent protein (GFP) reporter gene and transfecting fetus fibroblasts of normal livestock; sieving transgenic nuclear donor cells by using the GFP reporter gene and a flow cell sorter and introducing nuclear donor nuclei into denucleated oocytes of the livestock to obtain a reconstructed embryo; and immigrating the reconstructed embryo into the uterus of surrogate livestock to obtain an offspring, namely the FMDV-resistant RNAi heterozygote transgenic livestock. Heterozygote transgenic male livestock and heterozygote transgenic female livestock are normally mated or reproduced by artificial insemination so as to obtain homozygote transgenic livestock. The transgenic livestock prepared by the method can express the FMDV-resistant RNAi, obviously reduces the FMDV infection rate and has the capability of resisting foot-and-mouth disease (FMD).

Description

technical field [0001] The invention relates to an animal transgenic technology method for obtaining anti-foot-and-mouth disease virus RNAi transgenic livestock, and belongs to the field of cell engineering. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, febrile, highly contagious infection caused by foot-and-mouth disease virus (FMDV) in cattle, sheep, pigs and other cloven-hoofed animals. The World Organization for Animal Health lists it as the first class A severe infectious disease. Once an outbreak occurs, the infected and contact infected animals must be culled. In 2001, the outbreak of foot-and-mouth disease in the United Kingdom cost about 9 billion pounds, accounting for 1.1% of its GDP; in 2005, Asia I type FMD broke out in some parts of my country, and in early 2009, A type FMD occurred in many provinces, which not only caused huge losses. Direct economic losses, and seriously endanger the healthy development of animal husbandry and foreig...

AI Technical Summary

Problems solved by technology

FMDV mutates rapidly, has multiple serotypes, and there is no cross-protection between vaccines of different serotypes, so it is difficult to control and eradicate FMD by a single means of vaccine immunization

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