1. MicroRNA-Restricted Transgene Expression in the Retina.
- Author
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Karali, Marianthi, Manfredi, Anna, Puppo, Agostina, Marrocco, Elena, Gargiulo, Annagiusi, Allocca, Mariacarmela, Corte, Michele Della, Rossi, Settimio, Giunti, Massimo, Bacci, Maria Laura, Simonelli, Francesca, Surace, Enrico Maria, Banfi, Sandro, and Auricchio, Alberto
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MICRORNA ,TRANSGENE expression ,RETINA ,ADENO-associated virus ,GENETIC transformation ,TREATMENT effectiveness ,PHOTORECEPTORS - Abstract
Background: Gene transfer using adeno-associated viral (AAV) vectors has been successfully applied in the retina for the treatment of inherited retinal dystrophies. Recently, microRNAs have been exploited to fine-tune transgene expression improving therapeutic outcomes. Here we evaluated the ability of retinal-expressed microRNAs to restrict AAV-mediated transgene expression to specific retinal cell types that represent the main targets of common inherited blinding conditions. Methodology/Principal Findings: To this end, we generated AAV2/5 vectors expressing EGFP and containing four tandem copies of miR-124 or miR-204 complementary sequences in the 39UTR of the transgene expression cassette. These vectors were administered subretinally to adult C57BL/6 mice and Large White pigs. Our results demonstrate that miR-124 and miR- 204 target sequences can efficiently restrict AAV2/5-mediated transgene expression to retinal pigment epithelium and photoreceptors, respectively, in mice and pigs. Interestingly, transgene restriction was observed at low vector doses relevant to therapy. Conclusions: We conclude that microRNA-mediated regulation of transgene expression can be applied in the retina to either restrict to a specific cell type the robust expression obtained using ubiquitous promoters or to provide an additional layer of gene expression regulation when using cell-specific promoters. [ABSTRACT FROM AUTHOR]
- Published
- 2011
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