Molecular and Cellular and Genomics Biomedicine Research Group, Instituto de Investigación Sanitaria La Fe (IIS La Fe), 46026 Valencia, Spain.
Rare Diseases Joint Unit, CIPF-IIS La Fe, 46026 Valencia, Spain.
Genes (Basel). 2020 Apr 27;11(5):473. doi: 10.3390/genes11050473.
Inherited retinal dystrophies are an assorted group of rare diseases that collectively account for the major cause of visual impairment of genetic origin worldwide. Besides clinically, these vision loss disorders present a high genetic and allelic heterogeneity. To date, over 250 genes have been associated to retinal dystrophies with reported causative variants of every nature (nonsense, missense, frameshift, splice-site, large rearrangements, and so forth). Except for a fistful of mutations, most of them are private and affect one or few families, making it a challenge to ratify the newly identified candidate genes or the pathogenicity of dubious variants in disease-associated loci. A recurrent option involves altering the gene in in vitro or in vivo systems to contrast the resulting phenotype and molecular imprint. To validate specific mutations, the process must rely on simulating the precise genetic change, which, until recently, proved to be a difficult endeavor. The rise of the CRISPR/Cas9 technology and its adaptation for genetic engineering now offers a resourceful suite of tools to alleviate the process of functional studies. Here we review the implementation of these RNA-programmable Cas9 nucleases in culture-based and animal models to elucidate the role of novel genes and variants in retinal dystrophies.
遗传性视网膜病变是一组罕见疾病，它们共同构成了全球遗传性视力损害的主要原因。除了临床方面，这些视力丧失疾病还具有高度的遗传和等位基因异质性。迄今为止，已有超过 250 个基因与视网膜病变相关，报道的致病变异具有各种性质（无义、错义、移码、剪接位点、大片段重排等）。除了少数突变外，大多数都是罕见的，只影响一个或少数几个家族，这使得验证新鉴定的候选基因或疾病相关基因座中可疑变异的致病性具有挑战性。一种常见的方法是在体外或体内系统中改变基因，以对比产生的表型和分子印记。为了验证特定的突变，该过程必须依赖于模拟精确的遗传变化，而直到最近，这一过程一直具有挑战性。CRISPR/Cas9 技术的出现及其在基因工程中的适应性，为功能研究提供了一系列富有创意的工具。在这里，我们综述了这些 RNA 可编程 Cas9 核酸酶在基于培养和动物模型中的应用，以阐明在视网膜病变中新型基因和变异的作用。
