【Abstract】 Objective The objective of the study was to induce liver cancer formation in mice by stable overexpression of the protooncogene MYC using the Sleeping Beauty (SB) transposon and construct a recombinant transgene vector capable of knocking down the targeted gene PEX2 at the mouse level based on the SB transposon pT3EF1αcMyc. Methods Hydrodynamic tail vein injection (HTVI) and SB transposition systems were used to introduce the transposon plasmid carrying the target gene MYC into mouse liver to induce hepatoma formation. Using the vector as a template, U6shNC and U6shPEX2 sequences were amplified, and the target vectors pT3EF1αcMycshNC and pT3EF1αcMycshPEX2 were obtained through T4 ligation. Results Experimental mice successfully induced liver cancer. After identification by enzyme digestion and sequencing, the vectors were constructed correctly. Cell transfection confirmed that the experimental vector pT3EF1αcMycshPEX2 could reduce the expression of PEX2 in Hepa16 at the RNA level. Conclusion Overexpression of myc with HTVI technology and SB system successfully induces hepatoma formation in mice and obtains a transgenic vector capable of knocking down PEX2 gene based on the Sleeping Beauty transposon in mice.