Department of Ophthalmology, Institute of Biophysics, CCS, UFRJ, Rio de Janeiro, 21941-902, Brazil.
Adv Exp Med Biol. 2012;723:215-23. doi: 10.1007/978-1-4614-0631-0_29.
Small interfering RNA (siRNA) is a promising tool for the treatment of dominant diseases. Autosomal dominant eye disease like retinitis pigmentosa, are a leading cause of blindness. Mutations in lead to the degeneration of photoreceptors and are associated with several autosomal retinal diseases. Our goal is to develop a gene therapy for mutations. We describe a siRNA based mutation-independent approach, targeting in which levels of endogenous mutant and wild-type mRNA were reduced, and a siRNA-resistant version of gene was supplied simultaneously. siRNAs and resistant were delivered to the photoreceptors by recombinant adeno-associated virus (rAAV) vector through subretinal injections. The retinal phenotype was examined, both structurally and functionally at different time points after rAAV delivery. We demonstrate suppression of transcript by up to 50% with concomitant expression of replacement transcript in the retina of mice . These results validate the concept of suppression of and replacement strategies of gene therapy with rAAV vectors containing siRNA.
小干扰 RNA（siRNA）是治疗显性疾病的一种很有前途的工具。像色素性视网膜炎这样的常染色体显性眼病是导致失明的主要原因。突变导致光感受器退化，与几种常染色体视网膜疾病有关。我们的目标是开发针对突变的基因治疗方法。我们描述了一种基于 siRNA 的突变非依赖性方法，靶向，其中内源性突变型和野生型 mRNA 的水平降低，同时提供了一种 siRNA 抗性的基因版本。siRNA 和抗性基因通过重组腺相关病毒（rAAV）载体通过视网膜下注射递送至光感受器。在 rAAV 递送后的不同时间点，从结构和功能上检查视网膜表型。我们证明了在小鼠的视网膜中，通过高达 50%的抑制，同时伴随着替代转录物的表达。这些结果验证了 rAAV 载体中包含 siRNA 的抑制和基因治疗替代策略的概念。
