Screening And CeRNA Network Construction Of MiRNAs Related To Unsaturated Fatty Acid Synthesis In Bovine Adipocytes By Interference ACSL1 Gene

Beef is one of the main meat products,with high protein,low fat and rich vitamin content,is a kind of meat products favored by domestic and foreign markets.The content of unsaturated fatty acids in beef not only affects the flavor,but also improves the nutritional value of the meat.Therefore,it is important to elucidate the regulatory mechanism of unsaturated fatty acid synthesis in beef and further increase the content of unsaturated fatty acids in beef to improve beef quality.Long-chain lipoacyl Co A synthetase 1(ACSL1)plays a key role in the activation,transport,synthesis,and degradation of fatty acids.Previous studies in our group found that the expression of ACSL1 gene affected the composition of unsaturated fatty acids in bovine adipocytes by interfering and overexpressing ACSL1 gene in bovine adipocytes,and was a key gene for regulating the synthesis of unsaturated fatty acids in bovine adipocytes.In addition,circ RNA transcriptome sequencing that interfered with the ACSL1 gene in bovine adipocytes found that the relevant circ RNAs acted on the synthesis of unsaturated fatty acids in bovine adipocytes.In this study,small RNA transcriptome sequencing was performed by interfering with the ACSL1 gene in bovine adipocytes,screening and identifying the key miRNAs involved in regulating the synthesis of unsaturated fatty acids(UFAs)in bovine adipocytes,and combining circ RNAs sequencing results to construct circ RNA-miRNA-m RNA ce RNA interaction network.The main findings are as follows:1.RNA-Seq technology was used to screen for miRNAs related to the synthesis of unsaturated fatty acids.28 differentially expressed miRNAs(P-value < 0.05 and |fold change| > 2)were screened and the differentially expressed miRNAs were mainly involved in MAPK signaling pathways,metabolic pathways,fatty acid metabolism,fatty acid extension,etc.,signaling pathways related to the synthesis of unsaturated fatty acids.In the miRNA-m RNA interaction network,novel-m0035-5p — ACSL1,novel-m0035-5p —ELOVL4,miR-9-x — ACSL1,bta-miR-677 — ACSL1,miR-129-x — ELOVL4 and bta-miR-485—FADS2 targeting pairs were screened.2.The key miRNAs novel-m0035-5p and bta-miR-677,which act on the synthesis of unsaturated fatty acids,are highly conserved in mammals such as cattle and sheep.Combined analysis of bioinformatics prediction software found that bta-miR-677 and novel-m0035-5p targeted 739 and 1276 functional genes,respectively.These target genes are enriched in important biological processes and signaling pathways such as metabolic pathways,peroxisomes,and MAPK signaling pathways(P-value < 0.05).The time-series expression results showed that novel-m0035-5p and bta-miR-677 were expressed at the highest levels on days 0 and 4 of bovine adipocyte differentiation,respectively.3.The results of the miRNA-m RNA interaction network were further combined circ RNAs sequencing results showed that bta-miR-677 and bta-miR-485 were selected as important miRNAs,and two key ce RNA signal axes,novel＿circ＿0004161—bta-miR-677—ACSL1 and novel＿circ＿0001507—bta-miR-485—FADS2 were further screened.In summary,this study used RNA-seq technology,real-time PCR and bioinformatics analysis to screen miRNAs that may be involved in UFAs synthesis by interfering with and not interfering with the ACSL1 gene of bovine adipocytes,and constructed ce RNA regulatory network of circ RNA-miRNA-m RNA.The results provide a theoretical basis for further exploring the molecular mechanism of miRNAs regulating the synthesis of unsaturated fatty acids in bovine adipocytes.