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A Quantitative Dot Blot Assay for AAV Titration and Its Use for Functional Assessment of the Adeno-associated Virus Assembly-activating Proteins
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Structural characterization of an envelope-associated adeno-associated virus type 2 capsid

Joshua A Hull1, Mario Mietzsch1, Paul Chipman1

  • 1Department of Biochemistry and Molecular Biology, College of Medicine, Center for Structural Biology, McKnight Brain Institute, University of Florida, Gainesville, FL, 32610-0245, USA.

Virology
|October 12, 2021
PubMed

Insights

Envelope-associated adeno-associated virus (AAV) capsids show no structural differences from non-enveloped AAV. The MAAP protein may enhance exosome encapsulation, potentially offering therapeutic advantages.

Area of Science:

  • Virology
  • Structural Biology
  • Biotechnology

Background:

  • Adeno-associated viruses (AAV) are typically non-enveloped ssDNA viruses.
  • Exosome-associated AAV (EA-AAV) have been observed, with MAAP suggested in their formation.
  • Understanding EA-AAV structure is crucial for potential therapeutic applications.

Purpose of the Study:

  • To determine the cryo-EM structure of EA-AAV capsids.
  • To investigate the structural characteristics of EA-AAV compared to non-enveloped AAV.
  • To explore the role of MAAP in EA-AAV formation and potential therapeutic implications.

Main Methods:

  • Production of homogeneous EA-AAV capsids using the Sf9 baculoviral expression system.
  • Cryo-electron microscopy (cryo-EM) for structural determination.
  • High-resolution (3.14 Å) 3D reconstruction of the EA-AAV capsid.

Main Results:

  • The cryo-EM structure of AAV2 EA-AAV capsids was determined at 3.14 Å resolution.
  • EA-AAV capsids showed no significant structural variations compared to non-enveloped AAV capsids.
  • The Sf9 expression system suggests MAAP may enhance AAV encapsulation within exosomes.

Conclusions:

  • EA-AAV capsids are structurally similar to non-enveloped AAV.
  • MAAP may play a role in facilitating AAV encapsulation into exosomes.
  • EA-AAV may offer therapeutic benefits, including immune evasion and improved infectivity.
Keywords:
EA-AAV capsidEnveloped-AAVExo-AAVGene therapyMAAPSf9 productionStructure

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