Site-Specific Post-translational Surface Modification of Adeno-Associated Virus Vectors Using Leucine Zippers

ACS Synth Biol. 2020 Mar 20;9(3):461-467. doi: 10.1021/acssynbio.9b00341. Epub 2020 Feb 18.


Adeno-associated virus (AAV) is widely favored as a gene therapy vector, tested in over 200 clinical trials internationally. To improve targeted delivery a variety of genetic capsid modifications, such as insertion of targeting proteins/peptides into the capsid shell, have been explored with some success but larger insertions often have unpredictable deleterious impacts on capsid formation and gene delivery. Here, we demonstrate a modular platform for the integration of exogenous peptides and proteins onto the AAV capsid post-translationally while preserving vector functionality. We decorated the AAV capsid with leucine-zipper coiled-coil binding motifs that exhibit specific noncovalent heterodimerization. AAV capsids successfully display hexahistidine tagged-peptides using this approach, as demonstrated through nickel column affinity. This protein display platform may facilitate the incorporation of biological moieties on the AAV surface, expanding possibilities for vector enhancement and engineering.

Keywords: adeno-associated virus; gene delivery; protein engineering; synthetic virology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • CHO Cells
  • Capsid / metabolism
  • Capsid Proteins / genetics
  • Capsid Proteins / metabolism
  • Cricetulus
  • Dependovirus / genetics*
  • Genetic Engineering / methods*
  • Genetic Vectors / genetics*
  • Genetic Vectors / metabolism
  • Histidine / genetics
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Leucine Zippers / genetics*
  • Protein Processing, Post-Translational
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transduction, Genetic


  • Capsid Proteins
  • Recombinant Proteins
  • Histidine