Effective and Accurate Gene Silencing by a Recombinant AAV-Compatible MicroRNA Scaffold

Mol Ther. 2020 Feb 5;28(2):422-430. doi: 10.1016/j.ymthe.2019.11.018. Epub 2019 Nov 27.


Short hairpin RNAs that are delivered by recombinant adeno-associated virus (rAAV) have the potential to elicit long-term RNAi therapy for human disease. However, the discovery that short hairpin sequences can cause truncation of the rAAV genome calls into question the efficiency and gene-silencing specificity of this strategy in humans. Here, we report that embedding the guide strand of a small silencing RNA into an artificial microRNA (miRNA) scaffold derived from mouse miRNA-33 ensures rAAV genomic integrity and reduces off-targeting by 10-fold, while maintaining effective in vivo target gene repression in mice.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Dependovirus / genetics*
  • Gene Silencing*
  • Genetic Vectors / genetics*
  • Genome, Viral
  • Humans
  • Mice
  • MicroRNAs / genetics*
  • Nucleic Acid Conformation
  • RNA Interference
  • RNA Stability
  • RNA, Small Interfering / genetics
  • RNA, Viral


  • MIRN33a microRNA, human
  • MicroRNAs
  • RNA, Small Interfering
  • RNA, Viral