Optimization of AAV expression cassettes to improve packaging capacity and transgene expression in neurons

Mol Brain. 2014 Mar 11;7:17. doi: 10.1186/1756-6606-7-17.

Abstract

Adeno-associated virus (AAV) vectors can deliver transgenes to diverse cell types and are therefore useful for basic research and gene therapy. Although AAV has many advantages over other viral vectors, its relatively small packaging capacity limits its use for delivering large genes. The available transgene size is further limited by the existence of additional elements in the expression cassette without which the gene expression level becomes much lower. By using alternative combinations of shorter elements, we generated a series of AAV expression cassettes and systematically evaluated their expression efficiency in neurons to maximize the transgene size available within the AAV packaging capacity while not compromising the transgene expression. We found that the newly developed smaller expression cassette shows comparable expression efficiency with an efficient vector generally used for strong gene expression. This new expression cassette will allow us to package larger transgenes without compromising expression efficiency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / metabolism
  • Dependovirus / genetics
  • Dependovirus / physiology*
  • Gene Expression*
  • Genetic Techniques*
  • Genetic Vectors / metabolism*
  • HEK293 Cells
  • Humans
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neurons / metabolism*
  • Polyadenylation / genetics
  • Rats
  • Regulatory Sequences, Nucleic Acid / genetics
  • Sequence Deletion
  • Transgenes / genetics*
  • Virus Assembly / physiology*