Pseudo-virus and packaging method thereof and drug evaluation system

A pseudovirus and packaging technology, applied in the field of genetic engineering, can solve the problems of unable to make up for the lack of promoters, unable to drive the expression of wild-type S protein, etc., and achieve the effect of avoiding secondary infection

Active Publication Date: 2020-09-15
中吉当康(北京)基因技术有限公司
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Problems solved by technology

However, in 2004, when scientists packaged the SARS-CoV pseudovirus system, they found that in the packaging cells, the CMV promoter could not drive t

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  • Pseudo-virus and packaging method thereof and drug evaluation system
  • Pseudo-virus and packaging method thereof and drug evaluation system
  • Pseudo-virus and packaging method thereof and drug evaluation system

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Embodiment Construction

[0047] In order to better explain the present invention and facilitate understanding, the present invention will be described in detail below through specific embodiments in conjunction with the accompanying drawings.

[0048] The main innovation of the present invention is: when packaging pseudoviruses, CBA is selected as the promoter to drive the expression of the envelope protein Spike, and the Spike DNA sequence optimized for human expression is used to replace the wild-type Spike DNA sequence, thereby obtaining high-expression Lentiviral pseudoviruses of the S envelope protein. When constructing the new coronary pneumonia drug evaluation system of the present invention, a monoclonal 293T cell line that can overexpress and stably express the hACE2 receptor was also constructed to amplify the infection effect of the pseudovirus on cells, which is helpful for the evaluation of drug efficacy using the system , more accurate evaluation results can be obtained. In the process ...

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Abstract

The invention relates to pseudo-virus. The pseudo-virus comprises a plasmid for expressing envelope protein Spike, a reporter gene plasmid and a packaging helper plasmid, wherein the plasmid for expressing the envelope protein Spike contains a CBA promoter and a human optimized Spike expression sequence, and is used for driving the expression of the envelope protein Spike. The pseudo-virus disclosed by the invention can simulate the process of infecting cells by wild novel coronavirus, can be used for researching the relationship between virus and host cells, cloning a receptor of virus and replacing a traditional neutralization test, and is used for researching differential diagnosis of diseases, evaluation and screening of antiviral drugs, evaluation of vaccine immune effect and the like. The invention also relates to a packaging method of the pseudo-virus and a novel coronavirus virus drug evaluation system containing the pseudo-virus, wherein the pseudo-virus is used for infectingthe 293T monoclonal cell which overexpresses the human ACE2 gene under the condition of drug interference, and by detecting the expression quantity of a reporter gene in the pseudo-virus (such as fluorescence intensity detection), the effectiveness of the drug is preliminarily evaluated.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a pseudovirus, a packaging method thereof and a drug evaluation system. Background technique [0002] In the study of virus function, the most direct and efficient method is of course to manipulate wild-type live viruses. However, for highly pathogenic and infectious viruses such as the new coronavirus, the operation of wild-type live viruses needs to be carried out in biosafety protection (P3) laboratories above level 3. At present, there are only more than 40 P3 laboratories in China, and most of them Belongs to the CDC system, non-conventional scientific research purposes. In addition, in order to prevent biological hazards, the P3 laboratory has very strict protection conditions, which greatly reduces the operating efficiency. [0003] In order to be able to conduct research on the new coronavirus in the second-level biosafety laboratory, pseudovirus technology is a very ...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/50C12N15/867C12N5/10C12Q1/02
CPCC12N7/00C07K14/005C12N15/86G01N33/5008C12N2770/20022C12N2740/15043C12N2770/20021
Inventor 董文吉赵忠亮刘子瑾张艳君程谟斌李昌锋
Owner 中吉当康(北京)基因技术有限公司
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