Abstract: In order to predict the target genes of miR-2954 in Jingyuan chickens, analyze the expression differences of miR-2954 in different tissues of Jingyuan chickens, and to explore its regulatory mechanism in the growth and development of muscle tissues in Jingyuan chickens, bioinformatics methods were used to compare the mature sequences of miR-2954, and analyze its conserved properties. TargetScan, miRDB, miRmap online tools were used to predict the target genes, and then the target gene set was analyzed by GO functional annotation and KEGG signaling pathway enrichment, and the tissue expression level of miR-2954 was studied by real-time fluorescence quantitative PCR. The results showed that the miR-2954 sequence was highly conserved among birds, and the GO functional entries of the target genes mainly focused on the positive regulation of transcription by RNA polymerase Ⅱ, signal transduction, negative regulation of transcription by RNA polymerase Ⅱ, oxidation-reduction process, immune response, and G-protein coupled receptor signaling pathway. It existed in multiple components such as nucleus, cytoplasm, and exosomes, and also plays a role in molecular functions such as ATP binding, zinc ion binding, metal ion binding, protein binding and other molecular functions. The target genes were mainly enriched in signaling pathways such as cytokine-cytokine receptor interactions, regulation of actin cytoskeleton, cell adhesion molecules, AGE-RAGE signalling pathway in diabetic complications, insulin signalling pathway and adipocytokine signalling pathway and other signalling pathways. The relative expression level of miR-2954 in the lung and livers tissues of Jingyuan chickens was significantly higher than that in other tissues（P＜0.01）, and the relative expression of miR-2954 in lung, kidney, pectoral muscle and leg muscles of Jingyuan chickens was significantly higher than that in other organs（P＜0.01）. The results indicated that miR-2954 could affect the growth and development of Jingyuan chickens by regulating its target genes.