1. Targeting and silencing of rhodopsin by ectopic expression of the transcription factor KLF15.
- Author
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Botta S, de Prisco N, Marrocco E, Renda M, Sofia M, Curion F, Bacci ML, Ventrella D, Wilson C, Gesualdo C, Rossi S, Simonelli F, and Surace EM
- Subjects
- Animals, Dependovirus genetics, Ectopic Gene Expression, Female, Genetic Therapy methods, Genetic Vectors, Kruppel-Like Transcription Factors physiology, Mice, Transgenic, Mutation, Nuclear Proteins physiology, Retinal Rod Photoreceptor Cells metabolism, Retinitis Pigmentosa genetics, Retinitis Pigmentosa therapy, Rhodopsin metabolism, Swine, Gene Silencing, Gene Targeting methods, Kruppel-Like Transcription Factors genetics, Nuclear Proteins genetics, Rhodopsin genetics
- 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.
- Published
- 2017
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