Core Viewpoint - The article discusses the revolutionary AAVLINK technology, which overcomes the limitations of adeno-associated virus (AAV) in gene therapy by enabling the delivery of large genes, thus providing new treatment possibilities for genetic diseases such as autism and epilepsy [2][27]. Group 1: AAVLINK Technology Overview - AAVLINK stands for "AAV with translocation linkage," utilizing the Cre/lox system for DNA recombination, allowing large genes to be split into smaller segments and delivered via multiple AAVs [4]. - The technology enables precise reassembly of these gene segments within cells, leading to the expression of functional proteins [4]. Group 2: Advantages of AAVLINK - AAVLINK demonstrates significant advantages over existing methods for delivering large genes, such as protein splicing and RNA splicing, which often have low efficiency and produce many by-products [6]. - In experiments, AAVLINK achieved over 25 times higher efficiency in reconstructing fluorescent proteins compared to the intein method, and up to 245 times higher in triple vector delivery [12]. - AAVLINK produces minimal by-products, ensuring safety by avoiding truncated proteins that could interfere with normal functions [12]. Group 3: Application in Disease Models - AAVLINK was tested in two disease models: Phelan-McDermid syndrome (PMS) and Dravet syndrome, successfully delivering and reconstructing the SHANK3 and SCN1A genes, respectively, leading to improved behavioral and survival outcomes in mouse models [8][11]. Group 4: CRISPR Delivery Capability - AAVLINK can also deliver large CRISPR-Cas systems, enabling gene editing and regulation, which opens new avenues for treating genetic disorders, such as lowering cholesterol levels by editing the PCSK9 gene [11]. Group 5: Safety Enhancements and Resource Availability - The development of AAVLINK 2.0 addresses potential safety risks associated with the long-term presence of Cre enzyme in the body, paving the way for clinical applications [15][17]. - AAVLINK has established a resource library containing 198 large disease-related genes and 5 CRISPR tools, allowing researchers to accelerate therapy development [19][21].