Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype.
Mice homozygous for c.451C>T mutation in Cln1 gene recapitulate INCL phenotype.
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DOI：
10.1002/acn3.144
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发表时间：
2014-12
影响因子：
5.3
通讯作者：
Mukherjee, Anil B.
中科院分区：
文献类型：
--
作者：
Bouchelion, Ashleigh;Zhang, Zhongjian;Li, Yichao;Qian, Haohua;Mukherjee, Anil B.
无义突变占所有遗传疾病的5-70%。在美国，CLN 1/PPT 1基因的无义突变是>40%的婴儿神经元蜡样质脂褐质沉积症（INCL）患者的基础，INCL是一种破坏性的神经退行性溶酶体贮积病。我们试图产生一种可靠的携带在美国患者群体中发现的最常见的Ppt 1无义突变（c.451C>T）的INCL小鼠模型，以提供一个评估体内无义抑制因子的平台。我们使用靶向载体敲入C57胚胎干（ES）细胞中Ppt 1基因中的c.451C>T无义突变，其中LoxP位于Neo盒的侧翼，Neo盒通过电穿孔Cre从靶向ES细胞中去除。将两个独立靶向的ES克隆注射到胚泡中以产生同基因C57敲入小鼠，从而避免了广泛回交的必要性。通过DNA测序证实Ppt 1-KI小鼠的产生，其显示Ppt 1基因中存在c.451C>T突变。这些小鼠是可存活的和可生育的，尽管它们在中位年龄6个月时发展为痉挛（“抱握”表型）。自发荧光储存物质在整个大脑区域和内脏器官中积累。脑和脾的电子显微镜分析显示颗粒状嗜锇沉积物。神经元凋亡增加在大脑皮质中特别明显，异常的组织病理学和视网膜电图（ERG）分析证实了惊人的视网膜变性。运动协调和行为参数的进行性恶化持续到最终死亡。我们的研究结果表明，Ppt 1-KI小鼠可靠地重现INCL表型，为测试现有和新型无义抑制子的体内功效提供了平台。
Nonsense mutations account for 5–70% of all genetic disorders. In the United States, nonsense mutations in the CLN1/PPT1 gene underlie >40% of the patients with infantile neuronal ceroid lipofuscinosis (INCL), a devastating neurodegenerative lysosomal storage disease. We sought to generate a reliable mouse model of INCL carrying the most common Ppt1 nonsense mutation (c.451C>T) found in the United States patient population to provide a platform for evaluating nonsense suppressors in vivo. We knocked-in c.451C>T nonsense mutation in the Ppt1 gene in C57 embryonic stem (ES) cells using a targeting vector in which LoxP flanked the Neo cassette, which was removed from targeted ES cells by electroporating Cre. Two independently targeted ES clones were injected into blastocysts to generate syngenic C57 knock-in mice, obviating the necessity for extensive backcrossing. Generation of Ppt1-KI mice was confirmed by DNA sequencing, which showed the presence of c.451C>T mutation in the Ppt1 gene. These mice are viable and fertile, although they developed spasticity (a “clasping” phenotype) at a median age of 6 months. Autofluorescent storage materials accumulated throughout the brain regions and in visceral organs. Electron microscopic analysis of the brain and the spleen showed granular osmiophilic deposits. Increased neuronal apoptosis was particularly evident in cerebral cortex and abnormal histopathological and electroretinographic (ERG) analyses attested striking retinal degeneration. Progressive deterioration of motor coordination and behavioral parameters continued until eventual death. Our findings show that Ppt1-KI mice reliably recapitulate INCL phenotype providing a platform for testing the efficacy of existing and novel nonsense suppressors in vivo.
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影响因子：
4.2
作者：
HAMM, RJ;PIKE, BR;JENKINS, LW
通讯作者：
JENKINS, LW
影响因子：
14.9
作者：
BURKE, JF;MOGG, AE
通讯作者：
MOGG, AE
影响因子：
11.4
作者：
Hellsten, E;Vesa, J;Peltonen, L
通讯作者：
Peltonen, L
影响因子：
15.9
作者：
Chen, L;Adar, R;Deng, CX
通讯作者：
Deng, CX
DOI：
10.1073/pnas.251485198
发表时间：
2001-11-20
影响因子：
11.1
作者：
Gupta, P;Soyombo, AA;Hofmann, SL
通讯作者：
Hofmann, SL