首页> 美国卫生研究院文献>other >Unexpected CEP290 mRNA Splicing in a Humanized Knock-In Mouse Model for Leber Congenital Amaurosis
【2h】

Unexpected CEP290 mRNA Splicing in a Humanized Knock-In Mouse Model for Leber Congenital Amaurosis

机译:预期的CEP290 mRNA拼接在莱伯先天性人性化的人性化敲入小鼠模型中。

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务,本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文,但由于OA文献来源多样且变更频繁,仍可能出现获取不到、文献不完整或与标题不符等情况,如果获取不到我们将提供退款服务。请知悉。

摘要

Leber congenital amaurosis (LCA) is the most severe form of retinal dystrophy with an onset in the first year of life. The most frequent genetic cause of LCA, accounting for up to 15% of all LCA cases in Europe and North-America, is a mutation (c.2991+1655AG) in intron 26 of CEP290. This mutation generates a cryptic splice donor site resulting in the insertion of an aberrant exon (exon X) containing a premature stop codon to CEP290 mRNA. In order to study the pathophysiology of the intronic CEP290 mutation, we generated two humanized knock-in mouse models each carrying ~6.3 kb of the human CEP290 gene, either with or without the intronic mutation. Transcriptional characterization of these mouse models revealed an unexpected splice pattern of CEP290 mRNA, especially in the retina. In both models, a new cryptic exon (coined exon Y) was identified in ~5 to 12% of all Cep290 transcripts. This exon Y was expressed in all murine tissues analyzed but not detected in human retina or fibroblasts of LCA patients. In addition, exon x that is characteristic of LCA in humans, was expressed at only very low levels in the retina of the LCA mouse model. Western blot and immunohistochemical analyses did not reveal any differences between the two transgenic models and wild-type mice. Together, our results show clear differences in the recognition of splice sites between mice and humans, and emphasize that care is warranted when generating animal models for human genetic diseases caused by splice mutations.
机译:莱伯先天性黑ama病(LCA)是视网膜营养不良的最严重形式,发病于生命的第一年。 LCA的最常见遗传原因,占欧洲和北美所有LCA病例的15%,是CEP290内含子26中的突变(c.2991 + 1655AG)。这种突变产生了一个隐秘的剪接供体位点,导致一个异常的外显子(外显子X)插入到CEP290 mRNA中,该外显子含有一个过早的终止密码子。为了研究内含子CEP290突变的病理生理学,我们生成了两个人源化的敲入小鼠模型,每个模型均携带约6.3 kb的人CEP290基因,带有或不带有内含子突变。这些小鼠模型的转录特征揭示了CEP290 mRNA的意外剪接模式,尤其是在视网膜中。在这两种模型中,在所有Cep290转录本的约5%至12%中鉴定出新的隐秘外显子(硬币状外显子Y)。该外显子Y在所分析的所有鼠类组织中表达,但在人的视网膜或LCA患者的成纤维细胞中未检测到。此外,人类LCA特有的外显子x在LCA小鼠模型的视网膜中仅以非常低的水平表达。 Western印迹和免疫组织化学分析未发现两个转基因模型和野生型小鼠之间的任何差异。在一起,我们的结果表明小鼠和人类之间在剪接位点的识别上存在明显差异,并强调在生成由剪接突变引起的人类遗传疾病的动物模型时,必须格外小心。

著录项

相似文献

  • 外文文献
  • 专利
代理获取

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号