Dual AAV-mediated gene therapy restores hearing in a DFNB9 mouse model

Proc Natl Acad Sci U S A. 2019 Mar 5;116(10):4496-4501. doi: 10.1073/pnas.1817537116. Epub 2019 Feb 19.


Autosomal recessive genetic forms (DFNB) account for most cases of profound congenital deafness. Adeno-associated virus (AAV)-based gene therapy is a promising therapeutic option, but is limited by a potentially short therapeutic window and the constrained packaging capacity of the vector. We focus here on the otoferlin gene underlying DFNB9, one of the most frequent genetic forms of congenital deafness. We adopted a dual AAV approach using two different recombinant vectors, one containing the 5' and the other the 3' portions of otoferlin cDNA, which exceed the packaging capacity of the AAV when combined. A single delivery of the vector pair into the mature cochlea of Otof-/- mutant mice reconstituted the otoferlin cDNA coding sequence through recombination of the 5' and 3' cDNAs, leading to the durable restoration of otoferlin expression in transduced cells and a reversal of the deafness phenotype, raising hopes for future gene therapy trials in DFNB9 patients.

Keywords: DFNB9; deafness; dual AAV; gene therapy; otoferlin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Deafness / genetics
  • Deafness / therapy*
  • Dependovirus / genetics*
  • Disease Models, Animal
  • Genetic Therapy*
  • Genetic Vectors
  • Humans
  • Membrane Proteins / genetics*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic


  • Membrane Proteins
  • OTOF protein, human