Application of substance for inhibiting lnc-ALOX12 in preparation of influenza virus inhibitor

Abstract

The invention relates to the field of biological medicines, and discloses an application of a substance for inhibiting lnc-ALOX12 in preparation of an influenza virus inhibitor. According to the invention, the long-chain non-coding RNA lnc-ALOX12 is provided as a forward regulatory factor to participate in the replication process of the influenza virus for the first time. The IAV infection can be used for specifically promoting the expression of the lnc-ALOX12. The lnc-ALOX12 silencing can significantly inhibit transcription and replication of virus RNA and regulate and control influenza virus PB2 protein nucleation. And the lnc-ALOX12 participates in host adaptation mediated by the PB2 mutant. The invention reveals that lnc-ALOX12 plays an important role in an influenza virus replication process and cross-species transmission thereof, and provides a new basis for developing host lncRNA as an antiviral drug target.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the use of a substance that inhibits lnc-ALOX12 in the preparation of an influenza virus inhibitor. Background Art

[0002] Influenza viruses can cause acute respiratory infectious diseases, with influenza A virus (IAV) being the most threatening and prone to mutation. It is a highly contagious and pathogenic virus that is susceptible to humans and many animals. Over the past century, seasonal influenza and multiple influenza pandemics have posed serious threats and heavy losses to human life, health and the economy. Currently, the main means of preventing and treating influenza viruses in clinical practice are vaccines and anti-influenza drugs. However, due to the extremely strong ability of influenza viruses to recombine, mutate and drift antigenically, the development of new virus-specific vaccines has lagged behind and drug-resistant virus strains have continued to emerge, making the prevention and control of new viruses increasingly serious. Influenza virus is an important human pathogen that poses a major threat to global public health and the economy. The role of host lncRNAs in IAV infection has also received widespread attention.

[0003] Long noncoding RNA (lncRNA) refers to transcripts with a length of more than 200 nucleotides and low potential for encoding proteins. Compared with mRNA, lncRNA generally shows a more specific expression profile, with lower abundance and conservation. lncRNA plays a role in many important physiological processes such as genomic imprinting, immune response and nucleocytoplasmic transport.

[0004] The influenza virus RNA genome consists of eight segments of negative-sense single-stranded RNA, which are coated by nucleoprotein (NP) and heterologous polymerase complexes (PB1, PB2 and PA subunits) to form the viral ribonucleoprotein (vRNP) complex, which is an important functional unit for IAV genome transcription and replication. Evidence suggests that the PB1-PA dimer directly binds to RanBP5 and is transported into the nucleus via a non-classical pathway. PB2 and NP enter the nucleus via the classical pathway by directly interacting with importin-α / β1 dimers. Previous studies have shown that the adaptation of viral polymerase to the nuclear import mechanism is an important mechanism for the cross-species transmission of avian influenza viruses. Some adaptive mutations of viral polymerases, such as the PB2 subunit, especially positions 627 and 701, and N319K in NP, have been identified as being associated with the adaptation of avian IAVs to mammals, and these mutations enhance the binding of these proteins to importin-α. However, the exact regulatory mechanism of how host factors regulate the nuclear import of vRNPs remains to be clarified. Whether other cellular factors, especially lncRNA, are involved in the adaptation of the nuclear import mechanism by viral polymerase and mediate the interspecies transmission of influenza virus needs further exploration. Summary of the invention

[0005] The purpose of the present invention is to provide an application of a substance that inhibits lnc-ALOX12 in the preparation of an influenza virus inhibitor.

[0006] In order to achieve the purpose of the present invention, the present invention provides a human lncRNA whose function has not been reported, named lnc-ALOX12. Lnc-ALOX12 includes two transcripts, lnc-ALOX12-1 and lnc-ALOX12-2, which are transcribed from 14 exons of 6,891,943-6,954,939 nucleotides (NONHSAT145301.2 in the NONCODE v5 database (http: / / v5.noncode.org / index.php)) and 6,937,274-6,954,409 nucleotides (NONHSAT145307.2 in the NONCODE v5 database) of 17p13.1 of the hg38 genome, respectively.

[0007] In a first aspect, the present invention provides the use of a substance that inhibits lnc-ALOX12 in the preparation of an influenza virus inhibitor.

[0008] The lnc-ALOX12 is a long non-coding RNA from humans, and the sequences of its two transcripts are shown in SEQ ID NO: 1 (lnc-ALOX12-1) and SEQ ID NO: 2 (lnc-ALOX12-2).

[0009] In the present invention, the influenza virus is influenza A virus.

[0010] Preferably, the substance that inhibits lnc-ALOX12 is esilnc-ALOX12, and its target sequence acting on lnc-ALOX12 is shown in SEQ ID NO: 3.

[0011] In a second aspect, the present invention provides an influenza virus inhibitor, the active ingredient of which is the above-mentioned substance that inhibits lnc-ALOX12.

[0012] By means of the above technical solution, the present invention has at least the following advantages and beneficial effects: The present invention discloses for the first time that a long non-coding RNA lnc-ALOX12 participates in the replication process of influenza virus as a positive regulatory factor. IAV infection can specifically promote the expression of lnc-ALOX12. Silencing of lnc-ALOX12 can significantly inhibit the transcription and replication of viral RNA and regulate the nuclear entry of influenza virus PB2 protein. lnc-ALOX12 participates in host adaptation mediated by PB2 mutants. The present invention reveals that lnc-ALOX12 plays an important role in the replication process of influenza virus and its cross-species transmission, providing a new basis for the development of antiviral drugs targeting host lncRNA. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 In the preferred embodiment of the present invention, IAV infection induces the expression of lnc-ALOX12, and it is independent of IFN. A and B: HEK293T cells were infected with WSN for 24 hours (A) or infected at 0.5 moi for different times (B), and qRT-PCR was performed to detect lnc-ALOX12 expression (upper panel) and viral RNA levels (lower panel). C: The levels of lnc-ALOX12 (left panel) and viral RNA (right panel) in A549 cells infected with different influenza virus strains for 24 hours were measured by qRT-PCR. D: After A549 cells were treated with IFN-α (1,000 IU / mL) for different times, the levels of lnc-ALOX12 and MxA mRNA were measured by qRT-PCR. E: The levels of lnc-ALOX12 and IFN-β mRNA in 293T cells transfected with different amounts of poly(I:C) were detected by qRT-PCR.

[0014] AE data are Mean ± SD. p≤0.05, p≤0.01, NS, not significant (one-way ANOVA).

[0015] Figure 2Silencing lnc-ALOX12 inhibits IAV infection in a preferred embodiment of the present invention. AC: HEK293T-Gluc cells were reversely transfected with 10, 20, 40 nM esiEGFP or esilnc-ALOX12, and then infected with WSN (MOI = 0.5) for 24 hours. The knockdown efficiency of Lnc-ALOX12 was detected by qRT-PCR (A), and the infectivity of the virus was analyzed by measuring the Gluc reporter activity (B). The titer of the progeny virus was determined by the TCID50 method (C). D: The cell viability of HEK293T cells transfected with 10, 20, 40 nM esiEGFP or esilnc-ALOX12 was determined by the CCK8 assay after 48 hours. E: Different doses of lnc-ALOX12 were transfected into HEK293T-Gluc cells, and the infectivity of the virus was analyzed by measuring the Gluc reporter activity (left panel), and the expression level of lnc-ALOX12 was determined by qRT-PCR (right panel).

[0016] AE data are Mean ± SD. p≤0.05, p≤0.01, NS, not significant (two-tailed Student's t test (AD) and one-way ANOVA with Dunnett's post-test (E)).

[0017] Figure 3 Silencing of lnc-ALOX12 inhibits viral RNA transcription and replication in a preferred embodiment of the present invention. A: HEK293T cells were transfected with esiEGFP or esiEGFP and then infected with WSN (MOI=3). Immunofluorescence staining of viral NP (red) and cell nuclei (blue) was performed at different time points. B: In HEK293T cells transfected with esiEGFP or esiEGFP, RNP recombinant plasmids (PB1, PB2, PA, NP) and polI-luc reporter plasmids were transfected again. The effect of lnc-ALOX12 on the IAV replicon system was studied by measuring luciferase activity. C: HEK293T cells were transfected with esiEGFP or esiEGFP and then infected with WSN (MOI=0.5) for 24 hours, and the levels of viral NP mRNA / vRNA / cRNA were measured by qRT-PCR.

[0018] Data in B and C are Mean ± SD. p≤0.05, p≤0.01, NS, not significant (one-way ANOVA with Dunnett's post-test (B), two-tailed Student's t test (C)).

[0019] Figure 4 Silencing lnc-ALOX12 inhibits viral PB2 nuclear import in a preferred embodiment of the present invention. A: HEK293T cells were transfected with esiEGFP or different amounts of esilnc-ALOX12, and then co-transfected with plasmid DNA encoding vRNP subunits (PB1, PB2, PA, NP and Po1I-luc). Western analysis of viral PA, PB1 and PB2 protein levels. B: HEK293T cells were transfected with esiEGFP or esilnc-ALOX12, and then co-transfected with plasmid DNA encoding vRNP (PB1, PB2, PA, NP and Po1I-luc). After 6 hours of DMSO or MG132 treatment, the cells were subjected to nuclear-cytoplasmic separation, and Western analysis of the levels of viral PB2 in whole cells, cytoplasm and nucleus was performed. GAPDH protein was used as a cytoplasmic marker, and p84 protein was used as a nuclear marker. C: HEK293T cells were transfected with esiEGFP or esiLnc-ALOX12, and then co-transfected with plasmid DNA encoding PB1, PB2, PA, NP and Po1I-luc, and the cells were harvested after 6 hours of treatment with MG132. Cell lysates were immunoprecipitated and Western analyzed using control antibodies (IgG), anti-importin α1 or anti-importin β antibodies. D: PLA technology was used to analyze the effect of lnc-ALOX12 on the colocalization of PB2 and importin. Cell fields were randomly selected and the differences in red fluorescent spots were statistically analyzed (n=30, p≤0.001, two-tailed Student's t test).

[0020] Figure 5In the preferred embodiment of the present invention, nc-ALOX12 participates in host adaptation mediated by PB2 mutants. A: The activity of viral polymerase containing PB2 (WSN-PB2) and its mutants (avian-PB2 and mammal-PB2) was determined by luciferase activity. B: The binding of lnc-ALOX12 to PB2 (WSN-PB2) and its mutants (avian-PB2 and mammal-PB2) was detected by RIP technology. C: The effect of knocking down lnc-ALOX12 on the activity of viral polymerase containing PB2 (WSN-PB2) and its mutants (avian-PB2 and mammal-PB2) in human 293T cells. D: The effect of knocking down chicken chlnc-ALOX12 on the activity of viral polymerase containing PB2 (WSN-PB2) and its mutants (avian-PB2 and mammal-PB2) in chicken DF-1 cells. E: Effect of overexpression of chicken chlnc-ALOX12 on viral polymerase activity containing WSN-PB2 or avian-PB2 in human 293T cells. F: Effect of overexpression of chicken chlnc-ALOX12 on the interaction between importin and WSN-PB2 or avian-PB2 in human 293T cells was analyzed by co-immunoprecipitation. G: Effect of knockdown of chicken chlnc-ALOX12 on the interaction between importin and WSN-PB2 or avian-PB2 was analyzed by co-immunoprecipitation in chicken DF-1 cells.

[0021] (AE) Data are Mean ± SD. p≤0.05, p≤0.01, p≤0.001, NS, not significant (two-tailed Student's t test (CE)). DETAILED DESCRIPTION

[0022] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0023] The cell culture fluid used in the following examples is a DMEM medium containing 10% FBS. Influenza strain A / WSN / 33 is produced by 8-plasmid system virus rescue. Influenza virus A / WSN / 33 (H1N1) is produced by transfecting HEK293T cells with 8 plasmids (each plasmid encodes an IAV RNA fragment) and then co-culturing with MDCK according to the method described by Hoffmann et al. (Hoffmann, E., Neumann, G., Kawaoka, Y., Hobom, G., and Webster, RG (2000). A DNA transfection system for generation of influenza A virus from eight plasmids. Proc Natl Acad Sci USA 97, 6108-6113. 10.1073 / pnas.100133697.). Influenza viruses A / PR / 8 / 34 (H1N1), B / Beijinghaidian / 1386 / 2013 and B / Massachusetts / 02 / 2012 (provided by Dr. Shu Yuelong, China National Influenza Center), and A / Beijing / 30 / 95 (H3N2) (provided by Dr. Tan Wenjie, China Center for Disease Control and Prevention) were propagated in embryonated chicken eggs.

[0024] 293T, A549, MDCK, and DF-1 cells were purchased from the American Type Culture Collection (ATCC) (Manassas, VA). The 293T-Gluc cell line was constructed by transfection with the plasmid DNA pLenti6-Gluc to constitutively express the negative-strand RNA of the secreted luciferase (Gluc) gene. Gluc reporter gene activity can be used to evaluate the level of IAV infection. All cells were cultured in DMEM (Gibco, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS; Gibco) at 37°C in an incubator with 5% CO2, except for DF-1 cells, which were cultured at 39°C.

[0025] Example 1 IAV infection specifically induces lnc-ALOX12 expression We first evaluated the effect of IAV infection on the expression level of lnc-ALOX12. HEK293T cells were infected with IAV, and the levels of intracellular lnc-ALOX12 and viral RNA were measured by RT-qPCR 24 h later. Figure 1As shown in (A and B), the level of lnc-ALOX12 was upregulated with increasing viral RNA dose and time after infection. Similarly, lnc-ALOX12 was induced in cells infected with other IAVs (A / Beijing / 30 / 95 (H3N2) and A / PR / 8 / 34 (H1N1)), but not in cells infected with influenza B viruses (B / Beijinghaidian / 1386 / 2013 and B / Massachusetts / 02 / 2012) ( Figure 1 , C). Notably, we found that IFN-α ( Figure 1 , D) or poly(I:C) ( Figure 1 , E) Treatment can upregulate the positive control gene MxA ( Figure 1 , D) and IFN-β ( Figure 1 , E) mRNA levels, but did not affect the level of lnc-ALOX12 in cells. These data indicate that lnc-ALOX12 can be specifically induced by IAV infection, rather than being an immune response of the host to viral infection.

[0026] Example 2 Silencing of lnc-ALOX12 inhibits IAV infection This example uses two esiRNAs (endoribonuclease prepared siRNA, small interfering RNA prepared by endonuclease) named esiLnc-ALOX12 and esiEGFP. esiLnc-ALOX12 is an esiRNA used to silence the human lnc-ALOX12 gene. esiLnc-ALOX12 targets two transcripts of human lnc-ALOX12, lnc-ALOX12-1 and lnc-ALOX12-2. The target sequence (esiLnc-ALOX12 cDNA target sequence) of eSiLnc-ALOX12 acting on human lnc-ALOX12 is shown in SEQ ID NO: 3. esiLnc-ALOX12 is a product of Sigma, and its catalog number is EHNC006181.

[0027] esiEGFP is an esiRNA used as a negative control of esilnc-ALOX12, which targets EGFP and is an esiRNA used to silence the EGFP gene. esiEGFP is a product of Sigma, and its catalog number is EHUEGFP.

[0028] IAV promotes the expression of lnc-ALOX12. We studied whether the level of lnc-ALOX12 affects IAV replication. First, 293T cells were reversely transfected with esiEGFP or esilnc-ALOX12, and the transfection concentrations were set to 10nM, 20nM, and 40nM, respectively. After 24 hours, the cells were infected with influenza virus A / WSN / 33 (H1N1) with an MOI of 0.5. After 24 hours, the cells and supernatant were collected, and the real-time fluorescence quantitative PCR (RT-qPCR) technology was used to detect the knockdown effect of esiRNA on endogenous lnc-ALOX12 in cells. At the same time, the Gluc (secreted luciferase) activity and TCID50 value in the cell supernatant were measured to reflect the virus infection level and progeny virus titer, respectively. The results are shown in Figure 2. Figure 2 As shown, esilnc-ALOX12 (20 nM and 40 nM) significantly reduced the expression of lnc-ALOX12 ( Figure 2 , A). Compared with control cells, Gluc activity and viral titer in the cell supernatant decreased with the decrease of lnc-ALOX12 level ( Figure 2 , B and C). At the same time, the results of cytotoxicity experiments showed that compared with transfection with esiEGFP, transfection with esilnc-ALOX12 did not affect cell viability ( Figure 2 , D). Taken together, these results indicate that esiRNA-mediated silencing of lnc-ALOX12 inhibits IAV replication, and this inhibitory effect is not caused by cytotoxicity.

[0029] To further confirm the effect of lnc-ALOX12 on IAV replication, we transfected 293T-Gluc cells with DNA overexpression plasmids expressing lnc-ALOX12-1 and lnc-ALOX12-2, and then infected IAV (A / WSN / 33). The level of lnc-ALOX12 was determined by RT-qPCR, and the replication of IAV was analyzed by Gluc activity. The results showed that the overexpression of lnc-ALOX12 had no effect on the replication of IAV ( Figure 2 , E), indicating that the endogenous lnc-ALOX12 level induced by IAV infection is almost sufficient to meet the requirements of viral replication.

[0030] Example 3 lnc-ALOX12 gene silencing inhibits transcription and replication of viral RNA To determine the role of lnc-ALOX12 in the IAV replication cycle, we first used immunofluorescence to detect the effect of lnc-ALOX12 silencing on the nuclear import of viral RNPs after IAV infection. The results showed that within 6 hours after IAV infection of cells, lnc-ALOX12 silencing did not affect the nuclear import of viral RNPs ( Figure 3, A), indicating that lnc-ALOX12 is not involved in the early stages of IAV infection. However, in the IAV replicon system, as the amount of lnc-ALOX12 transfection increased, the IAV RNA polymerase activity decreased in a dose-dependent manner ( Figure 3 , B). Next, we used RT-qPCR to measure the levels of viral NP mRNA / vRNA / cRNA in 293T cells after lnc-ALOX12 gene silencing. The results showed that as the amount of lnc-ALOX12 transfection increased, the levels of the three NP transcripts (vRNA / mRNA / cRNA) gradually decreased ( Figure 3 , C). This indicates that lnc-ALOX12 plays a positive regulatory role in IAV transcription and replication.

[0031] Example 4 Silencing of lnc-ALOX12 inhibits viral PB2 nuclear import To further explore the mechanism by which lnc-ALOX12 silencing leads to reduced viral polymerase activity, we repeated the polymerase activity experiment. Western results showed that lnc-ALOX12 silencing significantly reduced the level of PB2 protein, while having little effect on the levels of PA and PB1 proteins ( Figure 4 , A). The nuclear-cytoplasmic separation experiment showed that compared with esiEGFP control cells ( Figure 4 , lane 6 in B) compared with lnc-ALOX12 silenced cells ( Figure 4 , Lane 8 in B) The nuclear PB2 level was significantly reduced, while treatment with the proteasome inhibitor MG132 could not significantly alleviate the effect of lnc-ALOX12 silencing on PB2 nuclear entry ( Figure 4 , B), indicating that lnc-ALOX12 silencing inhibited the nuclear import of PB2.

[0032] PB2 depends on the classical importin protein pathway, that is, nuclear import is achieved through direct interaction with importin-α / β1. Therefore, we tried to determine whether lnc-ALOX12 silencing would affect the interaction between PB2 and importin proteins. lnc-ALOX12 silenced or esiEGFP control cells were co-transfected with viral RNP subunit expression plasmids (PA, PB1, PB2 and NP) and viral mini-genome Po1I-luc. Immunoprecipitation was performed with anti-importin-α1 or anti-importin-β antibodies, and the level of PB2 in the precipitate was determined by Western blotting. The results showed that the binding of PB2 to importin-α1 and importin-β was significantly weakened after lnc-ALOX12 silencing ( Figure 4, C). Proximity ligation assay (PLA) is a powerful tool for in situ detection of molecular interactions with high specificity. The red fluorescent dots in the PLA image represent the binding of PB2 and importin. The fluorescence signal statistics of 30 cells were performed ( Figure 4 , D), lnc-ALOX12 silencing can significantly reduce the binding of the two, which is consistent with the above immunoprecipitation results. In summary, these results indicate that lnc-ALOX12 silencing inhibits the interaction between PB2 and importin protein and inhibits the nuclear import of PB2.

[0033] Example 5 lnc-ALOX12 is involved in PB2 mutant-mediated host adaptation The interaction between PB2 and importin protein plays an important role in the interspecies transmission of influenza viruses. Adaptive mutations in viral polymerases, especially PB2 subunits E627K and D701N, have been shown to be important host range determinants. Therefore, we further explored whether lnc-ALOX12 might be involved in host adaptation mediated by PB2 mutations. The WSN (H1N1) virus underwent an adaptive mutation at amino acid position 627 of PB2 (PB2 627K), but retained avian characteristics at amino acid position 701 of PB2 (PB2 701D). Based on this PB2 (WSN-PB2), we constructed two single mutants of the PB2 subunit, K627E and D701N, named avian-PB2 and mammal-PB2, respectively. As expected, the polymerase activity containing mammal-PB2 was increased by about 2.5 times, while the polymerase activity of avian-PB2 was significantly decreased ( Figure 5 , A), which is consistent with previous studies. Next, we used RIP technology to detect the binding ability of lnc-ALOX12 with PB2 and its mutants. The results showed that lnc-ALOX12 could bind to the wild-type PB2 protein of the WSN virus strain (WSN-PB2) and the D701N mutant PB2 protein (mammal-PB2), but almost did not bind to avian-PB2 ( Figure 5 , B). The same polymerase activity assay as above was performed in 293T cells transfected with si-ALOX12. The data showed that lnc-ALOX12 silencing significantly affected the polymerase activity of WSN-PB2 and mammal-PB2, but did not affect the polymerase activity of avian-PB2 ( Figure 5 , C). These results suggest a potential role for lnc-ALOX12 in the adaptation of avian influenza viruses to mammalian hosts.

[0034] To further verify the involvement of lnc-ALOX12 in the host adaptation process mediated by PB2 mutants, we first predicted the homologous lnc-ALOX12 in the avian genome (galGal6_dnarange=chr6:18475151-18481635) using the UCSC liftOver and AUGUSTUS tools, and obtained the sequence of chicken lnc-ALOX12 (chlnc-ALOX12). Knockdown of chlnc-ALOX12 in chicken DF-1 cells significantly reduced the polymerase activity of avian-PB2, but had relatively limited effects on the vRNP activity of mammalian characteristic WSN-PB2 and mammal-PB2 ( Figure 5 , D), indicating that chlnc-ALOX12 prefers to support the corresponding avian virus polymerase in chicken DF-1 cells. Similarly, overexpression of chlnc-ALOX12 can significantly enhance the polymerase activity of avian-PB2 in human 293T cells ( Figure 5 , E).

[0035] We further evaluated the effects of chlnc-ALOX12 overexpression and knockdown on the interaction between importin-α and WSN-PB2 or avian-PB2 by Co-IP assay in 293T and DF-1 cells. The results showed that overexpression of chlnc-ALOX12 in 293T cells did not affect the binding of mammalian WSN-PB2 to importin-α1, but enhanced the binding of avian-PB2 to importin-α1 ( Figure 5 , F). Similarly, in chicken DF-1 cells, knockdown of chlnc-ALOX12 reduced the level of avian-PB2 co-precipitated with importin-α3 but had no effect on the level of WSN-PB2 ( Figure 5 , G), indicating that the effect of lnc-ALOX12 on PB2 / importin interaction is species-specific.

[0036] In summary, the present invention discovered for the first time that lnc-ALOX12 species-specifically promotes the binding of PB2 to host importin, thereby ensuring the effective nuclear import of PB2 and viral RNA synthesis.

[0037] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. Application of substances inhibiting lnc-ALOX12 in the preparation of influenza virus inhibitors; The lnc-ALOX12 is a long non-coding RNA from humans, and the sequences of its two transcripts are shown in SEQ ID NO: 1 and SEQ ID NO:

2.

2. The use according to claim 1, characterized in that: The influenza virus is influenza A virus.

3. The use according to claim 1 or 2, characterized in that: The substance that inhibits lnc-ALOX12 is esilnc-ALOX12, and its target sequence acting on lnc-ALOX12 is shown in SEQ ID NO:

3.

4. An influenza virus inhibitor, characterized in that The active ingredient is a substance that inhibits lnc-ALOX12; The lnc-ALOX12 is a long non-coding RNA from humans, and the sequences of its two transcripts are shown in SEQ ID NO: 1 and SEQ ID NO:

2.

5. The influenza virus inhibitor according to claim 3, characterized in that The substance that inhibits lnc-ALOX12 is esilnc-ALOX12, and its target sequence acting on lnc-ALOX12 is shown in SEQ ID NO:3.

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