- vulnificus -infected wild- type and D56-transgenic tilapia using Illumina ’ s sequencing-by-synthesis approach. - Results: Comparative transcriptome analysis was performed on RNA-sequence profiles obtained from wild-type and D56- transgenic tilapia at 0, 6 and 24 h post-infection with V. - Conclusions: Transcriptome profiling and filtering for two-fold change variation showed that 3795 genes were upregulated and 1839 genes were downregulated in D56-transgenic tilapia. - The expression patterns of FA-related genes, inflammatory genes, antimicrobial peptide genes and immune responsive genes at and 48 h post-infection suggests these genes are involved in the enhanced resistance of D56 transgenic tilapia to V. - Keywords: Delta-6 desaturase and delta-5 desaturase (D56), D56 transgenic tilapia fish, RNA-seq, Fatty acid-associated genes, Immune responsive genes, Inflammatory genes. - In addition, the D56 transgenic tilapia exhibit altered gut microbial pro- files [15]. - vul- nificus-susceptible wild-type tilapia and D56 transgenic tilapia with enhanced resistance to the pathogen to re- veal the particular genes responsible for the resistance [15, 16]. - Wild-type and D56-transgenic tilapia were infected with V. - Transcriptome sequencing of six groups of samples produced a total of raw reads for wild-type and D56-transgenic tilapia. - Similarly and clean reads and 91.06% of raw reads) were obtained for D56-transgenic tilapia for 0, 6 and 24 hpi samples, re- spectively (Table 1). - vulnificus challenged wild-type and D56-transgenic tilapia (Supple- mentary Figure S1). - At the 0 h time-point, 3795 genes were upregulated and 1839 genes were downregulated in D56-transgenic tilapia (Fig. - We could recognize the relevance of DEG between wild-type and D56- transgenic tilapia at different time-points. - Fatty acid-associated genes are altered in D56-transgenic tilapia according to KEGG pathway analysis. - Gene enrichment analysis using the KEGG pathway database showed a total of 24, 30 and 33 pathways were affected in D56-transgenic tilapia at 0, 6, and 24 hpi, re- spectively (Table 2, 3 4). - Differences in the FA degrad- ation pathway were also observed between wild-type and D56-transgenic fish at 6 h post infection (Table 3), and the FA metabolism and FA degradation pathways were identified as differentially expressed at 24 hpi (Table 4).. - D56-transgenic tilapia exhibit altered immune-related gene expression in GO analysis. - WT-Liver-ctrl WT-Liver-6 h WT-Liver-24 h D56-Liver-ctrl D56-Liver-6 h D56-Liver-24 h RNA depletion of rRNA and organelle RNA was extracted from liver samples of wild-type and transgenic Tilapia fish expressing delta-6 desaturase plus delta-5 desaturase (D56) at 0, 6 and 24 h V. - D56-transgenic and wild-type tilapia at 0, 6 and 24 hpi with V. - Biological function-associated GO terms, such as defense response to bacterium, angiogenesis, im- mune response, antigen presenting and presentation and inflammatory response genes were also altered in D56- transgenic tilapia. - Taken together, the GO analysis revealed that inflammatory genes, chemokine-associated genes, cytokine-associated genes, immune-related genes and iron binding protein genes are differentially regulated in D56-transgenic and wild- type tilapia after V. - Since the KEGG analysis showed FA-associated path- ways are altered in D56-transgenic tilapia, the expression of selected FA pathway-associated genes was analyzed by real-time PCR at and 48 hpi with V. - Notably, ApoA4b was downregulated by D56- transgenic tilapia at 24 hpi (Fig. - 1 Delta 5 and Delta6 transgenic (D56) tilapia fish alters the transcriptome profile. - The comparative transcriptome data between wild-type / Delta 5 and Delta6 (D56) transgenic tilapia fish were mapped and differentially expressed genes were counted. - Wild-type and D56 transgenic tilapia fish liver RNA were compared in 0, 6 and 24 h post infection with V. - b Differentially expressed genes between wild-type and D56 transgenic tilapia fish at 0, 6 and 24 h V. - These results showed that FA metabolism-related genes are altered in transgenic tilapia upon V. - In addition to FA-associated genes, several inflammatory and immune responsive genes were altered in D56- transgenic tilapia according to the GO enrichment ana- lysis (Table 5, 6, 7). - vulnificus in wild-type and D56- transgenic tilapia liver.. - In D56-transgenic tilapia, the Complement C1q sub- component subunit B (C1qb) was upregulated at 6 hpi. - The AMPs also showed sig- nificant differences in expression between wild-type and D56-transgenic tilapia. - For the inflammatory factors, significant differences be- tween wild-type and D56-transgenic tilapia were de- tected for NF-κB2 at 12 and 24 hpi. - Tissue inhibitor Table 2 KEGG pathway enrichment analysis of wild-type and transgenic tilapia fish expressing delta-6 desaturase plus delta-5 desaturase (D56). - onl00564 Glycerophospholipid metabolism 8 0.0015. - onl00480 Glutathione metabolism 7 0.0019. - onl00340 Histidine metabolism 3 0.0133. - onl00052 Galactose metabolism 3 0.0389. - For inflammatory factors, cytokines and complement-related genes, we found that were sev- eral significant differences between wild-type and D56- transgenic tilapia whole blood. - Availability of whole genome se- quence and RNA sequence data in recent years has Table 3 KEGG pathway enrichment analysis of wild-type and transgenic tilapia fish expressing delta-6 desaturase plus delta-5 desaturase (D56) infected with V. - onl00620 Pyruvate metabolism 6 0.0001. - onl00561 Glycerolipid metabolism 7 0.0003. - onl00380 Tryptophan metabolism 6 0.0003. - onl00340 Histidine metabolism 4 0.0011. - onl00480 Glutathione metabolism 7 0.0013. - onl01200 Carbon metabolism 7 0.0131. - onl00360 Phenylalanine metabolism 2 0.0454. - The complex Bcl form has to be converted Table 4 KEGG pathway enrichment analysis of wild-type and transgenic tilapia fish expressing delta-6 desaturase plus delta-5 desaturase (D56) infected with V. - onl00830 Retinol metabolism 9 0.0006. - onl00640 Propanoate metabolism 5 0.0047. - onl00380 Tryptophan metabolism 6 0.011. - onl00052 Galactose metabolism 5 0.0121. - onl00620 Pyruvate metabolism 5 0.0185. - onl00360 Phenylalanine metabolism 3 0.0291. - onl01212 Fatty acid metabolism 6 0.0369. - Hence, we analyzed our transcriptome data with the KEGG pathway tool to search for major pathways altered in D56 transgenic tilapia compared to wild type. - expression by D56 transgenic tilapia fish compared to wild types.. - When tilapia Table 5 Gene ontology analysis of wild-type and transgenic tilapia fish expressing delta-6 desaturase plus delta-5 desaturase (D56). - D56-transgenic zebrafish and tilapia exhibit resistance to V. - A comparative analysis of liver transcriptomes of wild- type and D56-transgenic tilapia infected with V. - The KEGG pathway analysis showed several FA-associated pathways in wild-type and D56-transgenic tilapia groups at 0 hpi with V. - Several such pathways were also differ- entially activated between wild-type and D56-transgenic fish group 6 h and 24 h post infected with V. - The GO analysis showed that genes associated with in- flammation, chemokine synthesis, iron homeostasis and immune response are altered by D56 expression in Table 6 Gene ontology analysis of wild-type and transgenic tilapia fish expressing delta-6 desaturase plus delta-5 desaturase (D56) infected with V. - Table 7 Gene ontology analysis of wild-type and transgenic tilapia fish expressing delta-6 desaturase plus delta-5 desaturase (D56) infected with V. - vulnificus in D56-transgenic tilapia, sug- gesting the activation of these peptides was promoted by D56 to combat V. - The com- plement genes, C1qb, CFHR1 and CFD were also upreg- ulated in D56-transgenic tilapia after V. - It is also possible that the plasma anti- bacterial function is stimulated by D56 expression in transgenic tilapia [38, 39]. - The inflammation-associated gene expression included early upregulation of TLR-2 and TNF-α in D56-transgenic fish (Fig. - 2 Delta 5 and Delta 6 transgenic (D56) tilapia fish modulates the expression of Fatty acid associated gene expression. - Wild-type and D56 transgenic tilapia fish challenged with V. - Altogether, the FA-associated path- ways triggered in the D56-transgenic tilapia appear to regulate a variety of immune-related genes that may serve to enhance resistance to V. - We found that TLR-2, TLR-5 and ACKR4 were induced in the D56-transgenic line. - In this study, we measured the transcriptome-wide responses of wild- type and D56-transgenic tilapia to V. - Several pathways associated with inflammation, immune response, chemokine and cytochrome were specifically altered in D56-transgenic tilapia, as revealed by KEGG pathway and GO analysis of the wild-type and D56- transgenic tilapia at the baseline (0 hpi) and after infec- tion (6 and 24 hpi). - Wild-type and transgenic tilapia fish with dual expres- sion of delta-6 desaturase in liver and delta-5 desaturase. - 3 Delta 5 and Delta 6 transgenic (D56) tilapia fish alters the Complements gene expression. - 5 Delta 5 and Delta 6 transgenic (D56) tilapia fish modulates the pro-inflammatory cytokine gene expression. - Expression of D56 in transgenic Tilapia. - The delta 5 and delta 6 transgenic tilapia have been crossed and the offspring carrying D56 has been selected for the ex- periment. - 6 Delta 5 and Delta 6 transgenic (D56) tilapia fish alters PRDX1 , ACRK4 and TIMP2 gene expression. - 7 Delta 5 and Delta 6 transgenic (D56) tilapia fish modulates the pro-inflammatory cytokines and immune-related genes in whole blood.. - org/10.1186/s . - https://doi.org/10.3389/fmicb . - https://doi.org . - https://doi.. - https://doi.org/10.1 093/jn . - https://doi.org/10.1016/j.ejphar . - https://doi.org/10.1016/S . - https://doi.org/10.3390/. - https://doi.org/10.1016/j.fsi . - https://doi.org/10.1371/. - 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