Abstract: Background: Intramuscular fat (IMF) is one of the most important traits affecting meat quality such as tenderness, juiciness and flavor. Phosphotyrosine interaction domain containing 1 (PID1) gene has been reported as a candidate gene for fat deposition. Therefore, an overexpression of PID1 gene in pigs could be of great benefit to the swine industry and consumers. Materials and Methods: In this study, the PID1-transgenic pigs were produced by sperm mediated gene transfer and magnetic nanoparticles with high efficiency. Real-time fluorescence quantitative PCR and Southern blot methods were employed to identify PID1-transgenic pigs. The copy numbers of exogenous gene in transgenic pigs were estimated by Southern blot analysis. Results: Twenty four live piglets were born from the two sows by natural delivery. Eleven piglets were positive in the PCR amplifications (45.83%). These PCR-positive pigs were further identified by Southern-blot analysis, out of which 7 pigs were positive in both Southern blotting and PCR (29.16%), others were positive only in PCR. In addition, the PID1 expression levels in the liver and longissimus dorsi muscle tissue were higher than wild-type ones (p<0.05), whereas the levels in backfat and kidney were similar to wild-type ones in transgenic pigs. Conclusion: The results suggest that magnetic nanoparticles can be used to efficiently introduce a transgene into animals via sperm mediated gene transfer, the PID1-transgenic pig model could further serve to study obesity and the mechanism of IMF deposition.