China Animal Husbandry & Veterinary Medicine ›› 2025, Vol. 52 ›› Issue (7): 2981-2991.doi: 10.16431/j.cnki.1671-7236.2025.07.002

• Biotechnology • Previous Articles     Next Articles

Prediction of Biological Function of gga-miR-1574-5p and Verification of Its Targeting Relationship with G3BP2 Gene

PING Yuyu1,2, HUANG Xuan1, CAI Qingqing1, WANG Qiangzhou1, WANG Jiaxing2, BAI Hao1, CHEN Shihao1, CHANG Guobin1   

  1. 1. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
    2. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
  • Received:2024-09-26 Online:2025-07-05 Published:2025-07-01

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Abstract: 【Objective】 gga-miR-1574-5p is a miRNA associated with the cellular nutrient deprivation response.This study was aimed to analyze the function of gga-miR-1574-5p and identify its targeting relationship with G3BP2 gene,so as to provide a scientific basis for further analysis of the biological function of gga-miR-1574-5p. 【Method】 The mature sequence of gga-miR-1574-5p was obtained by online tool miRBase.Target genes of gga-miR-1574-5p were predicted using TargetScan database for GO function and KEGG pathway enrichment analysis,and the binding sites of G3BP2 gene and gga-miR-1574-5p were predicted.Dual luciferase reporter gene assay was used to verify the targeting relationship between gga-miR-1574-5p and G3BP2.Real-time quantitative PCR was used to detect the effect of gga-miR-1574-5p overexpression on G3BP2 gene expression.The effect of gga-miR-1574-5p on the expression of G3BP2 protein in chicken macrophages was detected by Western blotting. 【Result】 GO function enrichment results showed that the potential target genes of gga-miR-1574-5p were mainly enriched in cellular components such as nucleus and plasma membrane,biological processes such as regulation of transcription by RNA polymerase Ⅱ and positive regulation of transcription by RNA polymerase Ⅱ,and molecular functions such as protein binding and ATP binding.KEGG pathway enrichment results showed that the target genes were mainly concentrated in oocyte meiosis,TGF-beta signaling pathway,mitophagy-animal pathway,etc.A binding site between gga-miR-1574-5p seed region and the 3'-untranslated region (3'-UTR) of G3BP2 gene in Gallus gallus was detected in TargetScan database.Compared with G3BP2-3'-UTR wild-type plasmid and NC-mimics co-transfection group,the dual luciferase activity of G3BP2-3'-UTR wild-type plasmid and gga-miR-1574-5p mimics co-transfection group was extremely significantly decreased (P＜0.01).Real-time quantitative PCR results showed that compared with NC-mimics group,transfected with 20 nmol/L gga-miR-1574-5p mimics had no significant effect on the expression of G3BP2 gene (P＞0.05),but the expression of G3BP2 gene in 30 and 50 nmol/L gga-miR-1574-5p mimics groups were significantly or extremely significantly decreased (P＜0.05 or P＜0.01).Western blotting results showed that the expression of G3BP2 protein in 20,30 and 50 nmol/L gga-miR-1574-5p mimics groups were extremely significantly lower than that in NC-mimics group (P＜0.01). 【Conclusion】 gga-miR-1574-5p had a targeting relationship with G3BP2 gene,and gga-miR-1574-5p could inhibit the expression of G3BP2 gene and protein in chicken macrophages by specifically binding to G3BP2-3'-UTR.

Key words: gga-miR-1574-5p; target gene; G3BP2 gene; stress granules

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