Generation and Characterization of a genetic zebrafish model of SMA carrying the human SMN2 gene.
Generation and Characterization of a genetic zebrafish model of SMA carrying the human SMN2 gene.
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DOI：
10.1186/1750-1326-6-24
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发表时间：
2011-03-28
影响因子：
15.1
通讯作者：
Beattie CE
中科院分区：
文献类型：
--
作者：
Hao le T;Burghes AH;Beattie CE
人类疾病的动物模型是必不可少的，因为它们允许在细胞水平上分析疾病过程，并可以通过作为药物筛选和靶点验证的工具来推进治疗方法。在这里，我们报告了脊椎动物斑马鱼脊髓性肌萎缩症（SMA）的完整遗传模型的开发，以补充现有的斑马鱼，小鼠和无脊椎动物模型，并显示其用于测试改变SMN 2剪接的化合物的实用性。人类运动神经元疾病SMA是由运动神经元存活蛋白（SMN）的低水平（而不是完全缺乏）引起的。为了在斑马鱼中产生SMA的真实模型，我们已经产生了表达人SMN 2基因（hSMN 2）的转基因斑马鱼，其仅产生少量的全长SMN，并将其与smn-/-背景杂交。我们发现，人类SMN 2在斑马鱼中被剪接，就像在人类中一样，并产生低水平的SMN蛋白。此外，我们表明，反义寡核苷酸，增强正确的hSMN 2剪接增加全长hSMN RNA在这个模型中。当我们将这种转基因置于smn突变体背景下时，它拯救了smn突变体中发生的神经肌肉突触前SV 2缺陷，并增加了它们的存活率。我们已经产生了一个转基因鱼携带人类hSMN 2基因。该基因在鱼类中被剪接，就像在人类和小鼠中一样，这表明在这些脊椎动物中存在保守的剪接机制。此外，内含子剪接沉默子位点的反义靶向增加了从该转基因产生的全长SMN的量。将这种转基因移植到突变的smn鱼身上，挽救了突触前缺陷，提高了存活率。这种斑马鱼SMA模型具有人SMA的所有组分，因此可用于了解SMA中的运动神经元功能障碍，可用作增加全长SMN的药物或反义方法的体内测试，并可开发用于药物筛选。
Animal models of human diseases are essential as they allow analysis of the disease process at the cellular level and can advance therapeutics by serving as a tool for drug screening and target validation. Here we report the development of a complete genetic model of spinal muscular atrophy (SMA) in the vertebrate zebrafish to complement existing zebrafish, mouse, and invertebrate models and show its utility for testing compounds that alter SMN2 splicing. The human motoneuron disease SMA is caused by low levels, as opposed to a complete absence, of the survival motor neuron protein (SMN). To generate a true model of SMA in zebrafish, we have generated a transgenic zebrafish expressing the human SMN2 gene (hSMN2), which produces only a low amount of full-length SMN, and crossed this onto the smn-/- background. We show that human SMN2 is spliced in zebrafish as it is in humans and makes low levels of SMN protein. Moreover, we show that an antisense oligonucleotide that enhances correct hSMN2 splicing increases full-length hSMN RNA in this model. When we placed this transgene on the smn mutant background it rescued the neuromuscular presynaptic SV2 defect that occurs in smn mutants and increased their survival. We have generated a transgenic fish carrying the human hSMN2 gene. This gene is spliced in fish as it is in humans and mice suggesting a conserved splicing mechanism in these vertebrates. Moreover, antisense targeting of an intronic splicing silencer site increased the amount of full length SMN generated from this transgene. Having this transgene on the smn mutant fish rescued the presynaptic defect and increased survival. This model of zebrafish SMA has all of the components of human SMA and can thus be used to understand motoneuron dysfunction in SMA, can be used as an vivo test for drugs or antisense approaches that increase full-length SMN, and can be developed for drug screening.
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影响因子：
64.5
作者：
LEFEBVRE, S;BURGLEN, L;MELKI, J
通讯作者：
MELKI, J
影响因子：
3.5
作者：
McGovern, Vicki L.;Gavrilina, Tatiana O.;Burghes, Arthur H. M.
通讯作者：
Burghes, Arthur H. M.
影响因子：
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影响因子：
4.3
作者：
Ramesh, Tennore;Lyon, Alison N.;Beattie, Christine E.
通讯作者：
Beattie, Christine E.
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14.8
作者：
Rembold, Martina;Lahiri, Kajori;Wittbrodt, Joachim
通讯作者：
Wittbrodt, Joachim