Engineered AAV13 variants with enhanced transduction and confined spread

Area of Science:

• Neuroscience
• Gene Therapy
• Molecular Biology

Background:

• Precise targeting of the central nervous system (CNS) is vital for brain disease research and gene therapy.
• Adeno-associated virus 13 (AAV13) offers restricted diffusion for targeting small brain regions but has low gene expression.
• Enhancing AAV13 transduction efficiency is critical for effective CNS applications.

Purpose of the Study:

• To engineer modified adeno-associated virus 13 (AAV13) capsid proteins to improve transduction efficiency.
• To evaluate the infectivity and CNS targeting capabilities of novel AAV13 variants.

Main Methods:

• Constructed AAV13-YF by mutating surface tyrosine to phenylalanine.
• Inserted the 7m8 peptide into AAV13 capsid positions 587/588 and 585/586, creating AAV13-587-7m8 and AAV13-585-7m8.
• Assessed in vitro infectivity in HEK293T cells and in vivo CNS infection in C57BL/6 mice.

Main Results:

• AAV13-YF demonstrated superior in vitro infectivity compared to wild-type AAV13.
• AAV13-587-7m8 and AAV13-585-7m8 exhibited enhanced CNS infection capabilities in mice.
• AAV13-587-7m8 maintained a limited spread range within the CNS.

Conclusions:

• Engineered AAV13 variants, including AAV13-YF, AAV13-587-7m8, and AAV13-585-7m8, show enhanced transduction efficiency.
• These modified AAV13 vectors hold significant potential for precise gene delivery in neuroscience research and gene therapy for brain diseases.