Screening method of influenza virus related host gene mutants

Abstract

The invention discloses a screening method of influenza virus related host gene mutants, and belongs to the technical field of random mutation. In order to screen out host genes related to influenza viruses, the invention provides a screening method. The screening method comprises the following steps: 1) transfecting a human lung cancer cell strain with CAG-tTA plasmids to obtain a Tel-off cell strain of a transcription activator stably expressed and regulated by tetracycline; co-transfecting the Tel-off cell strain with a gene search vector and plasmids that effectively express transposase toobtain a whole-genome pulmonary epithelial cell gene mutation library; 2) infecting the mutation library obtained in the step 1) by using the H1N1-WSN influenza virus; and 3) extracting the genome DNA of the cell strain, resisting the H1N1-WSN influenza virus, obtained in the step 2), and carrying out sequencing by Splinkette PCR to obtain the influenza virus related host gene mutants. The screening method can be used for screening the influenza virus related host gene mutants.

Description

technical field [0001] The invention relates to a method for screening influenza virus-related host gene mutants, which belongs to the technical field of random mutations. Background technique [0002] Seasonal influenza is an acute respiratory infectious disease and a serious public health problem for all mankind. Its characteristics include easy outbreaks, easy spread, easy evolution, wide susceptible population, easy seasonal epidemics, and high incidence. Due to the high mutation rate of influenza virus and the recombination between viruses, it is difficult for the existing anti-influenza virus drugs and vaccines to effectively deal with the variable influenza virus. Compared with viral proteins, host factors involved in viral infection and expansion are less likely to mutate, and are important targets for intervening in viral replication and inhibiting host responses. Therefore, screening and identification of key host factors against influenza virus may be a breakthro...

AI Technical Summary

Problems solved by technology

RNA interference, cDNA library, CRISPR / Cas9 mutation, and chip technology need to know the sequence of the target gene in advance, which will narrow the scope of gene screening, and often miss some unknown genes and non-coding regions; high-throughput sequencing and chip technology There is no clear causal relationship between the genes and influenza virus infection. Many key host genes may only be up-regulated or down-regulated at a certain time. These static analyzes are likely to ignore these genes; in addition, gene chips or sequencing The data obtained by the technology is huge, and the expression of thousands of genes is different, which brings great difficulties to the follow-up analysis work

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