Structure of AAV-DJ, a retargeted gene therapy vector: Cryo-electron microscopy at 4.5 Å resolution

Thomas F. Lerch, Jason K. O'Donnell, Nancy L. Meyer, Qing Xie, Kenneth A. Taylor, Scott M. Stagg, Michael Chapman

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

AAV-DJ, a leading candidate vector for liver gene therapy, was created through random homologous recombination followed by directed evolution, selecting for in vivo liver tropism and resistance to in vitro immune neutralization. Here, the 4.5 Å resolution cryo-EM structure is determined for the engineered AAV vector, revealing structural features that illuminate its phenotype. The heparan sulfate receptor-binding site is little changed from AAV-2, and heparin-binding affinity is similar. A loop that is antigenic in other serotypes has a unique conformation in AAV-DJ that would conflict with the binding of an AAV-2 neutralizing monoclonal antibody. This is consistent with increased resistance to neutralization by human polyclonal sera, raising the possibility that changed tropism may be a secondary effect of altered immune interactions. The reconstruction exemplifies analysis of fine structural changes and the potential of cryo-EM, in favorable cases, to characterize mutant or ligand-bound complexes.

Original languageEnglish (US)
Pages (from-to)1310-1320
Number of pages11
JournalStructure
Volume20
Issue number8
DOIs
StatePublished - Aug 8 2012

Fingerprint

Cryoelectron Microscopy
Tropism
Genetic Therapy
Heparitin Sulfate
Liver
Homologous Recombination
Neutralizing Antibodies
Heparin
Binding Sites
Monoclonal Antibodies
Ligands
Phenotype
Serum
In Vitro Techniques
Serogroup

ASJC Scopus subject areas

  • Molecular Biology
  • Structural Biology

Cite this

Lerch, T. F., O'Donnell, J. K., Meyer, N. L., Xie, Q., Taylor, K. A., Stagg, S. M., & Chapman, M. (2012). Structure of AAV-DJ, a retargeted gene therapy vector: Cryo-electron microscopy at 4.5 Å resolution. Structure, 20(8), 1310-1320. https://doi.org/10.1016/j.str.2012.05.004

Structure of AAV-DJ, a retargeted gene therapy vector : Cryo-electron microscopy at 4.5 Å resolution. / Lerch, Thomas F.; O'Donnell, Jason K.; Meyer, Nancy L.; Xie, Qing; Taylor, Kenneth A.; Stagg, Scott M.; Chapman, Michael.

In: Structure, Vol. 20, No. 8, 08.08.2012, p. 1310-1320.

Research output: Contribution to journalArticle

Lerch, TF, O'Donnell, JK, Meyer, NL, Xie, Q, Taylor, KA, Stagg, SM & Chapman, M 2012, 'Structure of AAV-DJ, a retargeted gene therapy vector: Cryo-electron microscopy at 4.5 Å resolution', Structure, vol. 20, no. 8, pp. 1310-1320. https://doi.org/10.1016/j.str.2012.05.004
Lerch TF, O'Donnell JK, Meyer NL, Xie Q, Taylor KA, Stagg SM et al. Structure of AAV-DJ, a retargeted gene therapy vector: Cryo-electron microscopy at 4.5 Å resolution. Structure. 2012 Aug 8;20(8):1310-1320. https://doi.org/10.1016/j.str.2012.05.004
Lerch, Thomas F. ; O'Donnell, Jason K. ; Meyer, Nancy L. ; Xie, Qing ; Taylor, Kenneth A. ; Stagg, Scott M. ; Chapman, Michael. / Structure of AAV-DJ, a retargeted gene therapy vector : Cryo-electron microscopy at 4.5 Å resolution. In: Structure. 2012 ; Vol. 20, No. 8. pp. 1310-1320.
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