Transcriptomics Analysis Of Brain And Spleen Of Zebrafish With TRIM47 Knockout And Antiviral Study Of USP5

The immune system is an important system to perform immune responses and immune functions.Many proteins in the body will activate the immune system when stimulated by the outside world.TRIM family and USP family proteins play an important role in many processes of life.Tripartite motif protein（TRIM）is a conserved protein that plays an important role in many biological processes,such as apoptosis,cell cycle regulation,cell response to viruses,and inflammation.In this study,the spleen of immune organs and the highly expressed tissue brain in TRIM47carp were used as experimental materials.The transcriptomics analysis of the brain and spleen of TRIM47 knockout zebrafish was used to clarify the possible role of TRIM47 in zebrafish.To study the role and mechanism of USP5 protein in innate immunity.It provides a theoretical basis for further understanding of the natural immunity of teleost fish.The main contents are as follows:1. Transcriptomics analysis of brain and spleen of TRIM47 knockout zebrafishIn this study,CRISPR/Cas9 technology was used to knock out the TRIM47gene of zebrafish,and the brain and spleen of TRIM47^-/-（TRIM47 knockout）and WT（wild type）zebrafish were collected for RNA-seq analysis to identify differentially expressed genes（Difference Expression Genes,DEGs）.In this study,271 DEGs in the brain and spleen were identified in the knockout group compared to wild-type zebrafish.After a brief analysis of these DEGs,these DEGs were annotated as the KEGG pathway and subjected to GO enrichment analysis.Compared with the WT group,the DEGs in the TRIM47^-/-group were concentrated in the cell adhesion and glutamatergic synaptic signaling pathways;in the spleen,compared with the wild-type group,DEGs have changed in the complement and coagulation cascade signaling pathways.The genes related to the complement pathway in the brain and spleen were verified by q RT-PCR,which was consistent with the transcriptome data.These results indicate that TRIM47 plays an important biological role,especially in the innate immune function through the complement pathway.SVCV infection experiments were carried out on the TRIM47 knockout and wild-type zebrafish,and the survival rate of the zebrafish in the TRIM47 knockout group was lower.These findings provide a basis for further research on the function of TRIM members in innate immunity.2. USP5 anti-virus researchThe Ubiquitin-Specific Protease（USP）family is involved in many life processes.Anti-virus is one of the important functions,and activating interferon is an important anti-virus method.This study aims to clarify the antiviral mechanism of zebrafish ubiquitin-specific protease 5（USP5）.By analyzing the evolutionary tree and protein sequence homology of USP5,the location of USP5 in FHM cells was determined.Overexpression of USP5 can activate higher m RNA levels of IFNφ1 and ISGs,and promoter activity of IFNφ1 and IFN-stimulated response elements（ISRE）is also activated.The study found that USP5 can enhance the expression of RIG-I.After treatment with Poly（I:C）,it can stimulate a higher level of IFNφ1 compared to the empty vector.Viral infection and interference tests showed that USP5 inhibited SVCV replication in vitro.The results of this study show that USP5 can activate higher levels of interferon by increasing RIG-I protein levels,thereby achieving antiviral function,which also provides a certain theoretical study for fish antiviral.