Extracellular vesicle (EV)-based low-density lipoprotein receptor (Ldlr) mRNA delivery showed excellent therapeutic effects in treating familial hypercholesterolemia (FH). Nevertheless, the loading inefficiency of EV-based mRNA delivery presents a significant challenge. Recently, RNA-binding proteins (RBPs) have been fused to EV membrane proteins for selectively encapsulating targeted RNAs to promote loading efficiency. However, the strong interaction between therapeutic RNAs and RBPs prevents RNA release from endosomes to the cytosol in the recipient cells. In this study, an improved strategy was developed for efficient encapsulation ofLdlrmRNA into EVs in donor cells and controllable release in recipient cells.Methods:The MS2 bacteriophage coat protein (CD9-MCP) fusion protein,LdlrmRNA, and a customized MS2 containing RNA aptamer base-pair matched withLdlrmRNA were expressed in donor cells. Cells receiving the above therapeutic EVs were simultaneously treated with EVs containing “Ldlrreleaser” with a sequence similar to the recognition sites inLdlrmRNA. Therapeutic effects were analyzed inLdlr-/-mice receiving EV treatments via the tail vein.Results:In vitroexperiments demonstrated improved loading efficiency ofLdlrmRNA in EVs via MS2-MCP interaction. Treatment of “Ldlrreleaser” competitively interacted with MS2 aptamer with higher affinity and releasedLdlrmRNA from CD9-MCP for efficient translation. When the combinatory EVs were delivered into recipient hepatocytes, the robust LDLR expression afforded therapeutic benefits inLdlr-/-mice.Conclusion:We proposed an EV-based mRNA delivery strategy for enhanced encapsulation of therapeutic mRNAs in EVs and RNA release into the cytosol for translation in recipient cells with great potential for gene therapy.