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Receptor-guided AAV Tropism Engineering via MATCH.

Nolan Graham1, Satheesh Kumar1, Joseph Rainaldi1,2

  • 1Department of Bioengineering, University of California San Diego, CA USA.

Biorxiv : the Preprint Server for Biology
|April 10, 2026
PubMed
Summary
This summary is machine-generated.

Researchers developed MATCH, a method to control adeno-associated virus (AAV) tropism by conjugating targeting proteins. This enables precise gene delivery, enhancing viral targeting for therapeutic applications.

Keywords:
AAV capsid engineeringBlood-brain barrierImmune cell engineeringSpyTag/SpyCatcherTargeted gene deliveryTransferrin receptor (TfR1)

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Gene Therapy

Background:

  • Controlling viral tropism is crucial for effective gene delivery.
  • Adeno-associated viruses (AAVs) are promising gene therapy vectors but lack precise targeting.
  • Current methods for retargeting AAVs are often inefficient or lack control.

Purpose of the Study:

  • To develop a modular method for programmable retargeting of AAVs.
  • To enable site-specific covalent conjugation of targeting proteins to AAV capsids.
  • To engineer AAV vectors with enhanced tropism for specific cell types and tissues.

Main Methods:

  • Developed MATCH (Modulation of AAV Tropism through Conjugation to Homing proteins), a biochemical method using SpyTag/SpyCatcher system for protein conjugation.
  • Incorporated SpyTag peptide into AAV capsid loops for site-specific attachment of SpyCatcher-linked targeting proteins.
  • Utilized mosaic AAV-DJ and AAV9 capsids with controlled SpyTag incorporation for tunable ligand display.

Main Results:

  • MATCH-AAVs conjugated to anti-CD3 antibodies efficiently transduced human T cells in vitro.
  • TfR1-targeted MATCH-AAV9 showed up to an 84-fold increase in brain transduction in vivo compared to wild-type AAV9.
  • Demonstrated receptor-dependent transduction and blood-brain barrier crossing with TfR1-targeted vectors.
  • Developed a one-pot "Mix-and-MATCH" production strategy yielding functional, targeted AAVs.

Conclusions:

  • MATCH provides a versatile and generalizable synthetic-biology approach for rational AAV tropism engineering.
  • This method enables scalable production of custom AAV vectors for research and therapeutic applications.
  • Programmed retargeting of AAVs via covalent protein conjugation offers a powerful tool for gene therapy development.