New AAVLINK Method Enables Delivery of Large Therapeutic Genes

Delivery of therapeutic genes is essential for gene therapy. Adeno-associated viruses (AAVs) are a prime vector for carrying gene cargoes because of their superior gene segmentation flexibility and robust gene reconstitution efficiency. However, their limited packaging capacity is a major challenge for large gene transduction.

In a study published in Cell, Prof. LU Zhonghua’s team from the Shenzhen Institute of Advanced Technology of the Chinese Academy of Sciences, and collaborators from Peking University First Hospital developed the AAV with translocation LINKage (AAVLINK), which harnesses Cre/lox-mediated intermolecular DNA recombination to enable in vivo reassembly of large genes.

AAVLINK allows flexible gene segmentation design, achieves high-efficiency full-length gene reconstitution, and markedly reduces the production of aberrant truncated proteins compared with conventional approaches.

Using animal models, the researchers found that AAVLINK enabled robust expression of full-length Shank3 and significantly rescued autism-like behavioral phenotypes in Shank3-deficient mice. Similarly, AAVLINK-mediated delivery of the large epilepsy-associated gene SCN1A restored gene expression and alleviated seizure phenotypes in mutant mice. These findings provide strong evidence that AAVLINK supports functional delivery of large therapeutic genes in the nervous system.

Furthermore, the researchers developed AAVLINK 2.0 by incorporating a destabilized Cre recombinase. This design provides tighter temporal control of the recombination activity, reducing potential safety concerns while preserving high gene reconstitution efficiency.

Using the AAVLINK strategy, the researchers constructed a vector bank covering 193 large genes associated with inherited disorders, including autism and epilepsy, and validated the gene reconstitution capacity of all constructs. The vector bank also includes five CRISPR-based genetic tools, demonstrating the broad applicability of the AAVLINK platform.

The study proposes a strategy that enables the delivery of large gene cargoes using AAVs, thereby offering therapeutic possibilities for diseases previously considered inaccessible to AAV-based gene therapy.

Reference: Lin J, Lin Y, Liu N, et al. AAVLINK: A potent DNA-recombination method for large cargo delivery in gene therapy. Cell. 2026. doi: 10.1016/j.cell.2025.12.039

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