Targeting and silencing of rhodopsin by ectopic expression of the transcription factor KLF15

JCI Insight. 2017 Dec 21;2(24):e96560. doi: 10.1172/jci.insight.96560.

Abstract

The genome-wide activity of transcription factors (TFs) on multiple regulatory elements precludes their use as gene-specific regulators. Here we show that ectopic expression of a TF in a cell-specific context can be used to silence the expression of a specific gene as a therapeutic approach to regulate gene expression in human disease. We selected the TF Krüppel-like factor 15 (KLF15) based on its putative ability to recognize a specific DNA sequence motif present in the rhodopsin (RHO) promoter and its lack of expression in terminally differentiated rod photoreceptors (the RHO-expressing cells). Adeno-associated virus (AAV) vector-mediated ectopic expression of KLF15 in rod photoreceptors of pigs enables Rho silencing with limited genome-wide transcriptional perturbations. Suppression of a RHO mutant allele by KLF15 corrects the phenotype of a mouse model of retinitis pigmentosa with no observed toxicity. Cell-specific-context conditioning of TF activity may prove a novel mode for somatic gene-targeted manipulation.

Keywords: Neurodegeneration; Ophthalmology; Therapeutics; Transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Dependovirus / genetics
  • Ectopic Gene Expression
  • Female
  • Gene Silencing*
  • Gene Targeting / methods*
  • Genetic Therapy / methods
  • Genetic Vectors
  • Kruppel-Like Transcription Factors / genetics*
  • Kruppel-Like Transcription Factors / physiology
  • Mice, Transgenic
  • Mutation
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / physiology
  • Retinal Rod Photoreceptor Cells / metabolism
  • Retinitis Pigmentosa / genetics
  • Retinitis Pigmentosa / therapy
  • Rhodopsin / genetics*
  • Rhodopsin / metabolism
  • Swine

Substances

  • KLF15 protein, human
  • Kruppel-Like Transcription Factors
  • Nuclear Proteins
  • Rhodopsin