Triacylglycerol (TGA) is the primary component of intramuscular fat. Expression of diacylglyceryl transferase (
DGAT) determines the polyester differentiation ability of precursor adipocytes. The two
DGAT isoforms (
DGAT1 and
DGAT2) play different roles in TAG metabolism. This study investigates the
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Triacylglycerol (TGA) is the primary component of intramuscular fat. Expression of diacylglyceryl transferase (
DGAT) determines the polyester differentiation ability of precursor adipocytes. The two
DGAT isoforms (
DGAT1 and
DGAT2) play different roles in TAG metabolism. This study investigates the roles of
DGAT1 and
DGAT2 in signaling pathways related to differentiation and lipid metabolism in Yanbian bovine preadipocytes. sh-DGAT1 (sh-1), sh-DGAT2 (sh-2), and sh-DGAT1 + sh-DGAT2 (sh-1 + 2) were prepared using short interfering RNA (siRNA) interference technique targeting
DGAT1 and
DGAT2 genes and infected bovine preadipocytes. Molecular and transcriptomic techniques, including differentially expressed genes (DEGs) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis, were used to investigate the effects on the differentiation of Yanbian bovine preadipocytes. After interference with
DGAT1 and
DGAT2 genes, the contents of TAG and adiponectin were decreased. The TAG content in the sh-2 and sh-1 + 2 groups was significantly lower than that in the sh-NC group. RNA sequencing (RNA-seq) results showed 2070, 2242, and 2446 DEGs in the sh-1, sh-2, and sh-1 + 2 groups, respectively. The DEGs of the sh-2 group were mainly concentrated in the
PPAR,
AMPK, and
Wnt signaling pathways associated with adipocyte proliferation and differentiation. These results demonstrated that at the mRNA level,
DGAT2 plays a more important role in lipid metabolism than
DGAT1.
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