Production of VP3-only virus-like particles of Adeno-associated virus 2 in E. coli cells

Protein Expr Purif. 2024 Aug:220:106502. doi: 10.1016/j.pep.2024.106502. Epub 2024 May 15.

Abstract

Adeno-associated Virus (AAV) is a promising vector for gene therapy. However, few studies have focused on producing virus-like particles (VLPs) of AAV in cells, especially in E. coli. In this study, we describe a method to produce empty VP3-only VLPs of AAV2 in E. coli by co-expressing VP3 and assembly-activating protein (AAP) of AAV2. Although the yields of VLPs produced with our method were low, the VLPs were able to self-assemble in E. coli without the need of in vitro capsid assembly. The produced VLPs were characterized by immunological detection and transmission electron microscopy (TEM). In conclusion, this study demonstrated that capsid assembly of AAV2 is possible in E. coli, and E. coli may be a candidate system for production of VLPs of AAV.

Keywords: AAP; AAV production; AAV2; Capsid assembly; E. coli; Protein co-expression; VLP.

MeSH terms

  • Capsid Proteins* / biosynthesis
  • Capsid Proteins* / genetics
  • Capsid Proteins* / metabolism
  • Dependovirus* / genetics
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Genetic Vectors / chemistry
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Humans
  • Parvovirinae / genetics
  • Virion / genetics
  • Virion / metabolism
  • Virus Assembly

Substances

  • Capsid Proteins
  • VP3 protein, Dependovirus

Supplementary concepts

  • Adeno-associated virus-2