Aim PhiC31 integrase directs foreign DNA containing with attB integration into specific genomic DNA, incurring long-term and stable expression. This study analyzed the effect of LDLR expression mediated by phiC31 integrase on lipoprotein profile and atherosclerosis in family hypercholesterolaemia mice (LDLR－/－). Methods Male LDLR－/－ mice were tail vein co-injected with phiC31 integrase expression plasmid pCMV-int and plasmid pcDNA 3.1-TBG-LDLR-attB carried with LDLR gene and attB sequence. Mice co-injected with pCMV-int and pcDNA3.1-TBG-LDLR lacking attB sequence were used as control. Results LDLR directed by thyroxine binding globulin promoter and mediated by phiC31 integrase maintained higher and persistent expression in H22 cells and specifically in livers from mice injected with pcDNA3.1-TBG-LDLR-attB. The expression reduced serum low density lipoprotein cholesterol (LDLC) levels up to 35%, and resulted in decreases by 19% of plaque size/lumen area in cross section at ascending aorta compared with mice co-injected with pCMV-int and pcDNA3.1-TBG-LDLR. Conclusion The expression of LDLR mediated by phiC31 integrase improved lipoprotein profile and ameliorated atherosclerosis in LDLR－/－ mice.