Detection of human survival motor neuron (SMN) protein in mice containing the SMN2 transgene: applicability to preclinical therapy development for spinal muscular atrophy.
Detection of human survival motor neuron (SMN) protein in mice containing the SMN2 transgene: applicability to preclinical therapy development for spinal muscular atrophy.
复制标题
DOI：
10.1016/j.jneumeth.2008.07.024
复制
发表时间：
2008-10-30
影响因子：
3
通讯作者：
Lorson, Christian L.
中科院分区：
文献类型：
--
作者：
Mattis, Virginia B.;Butchbach, Matthew E. R.;Lorson, Christian L.
关键词：
脊髓性肌萎缩症（SMA）是婴儿死亡的主要遗传原因，其原因是脊髓运动神经元的丧失导致骨骼肌萎缩。SMA是由生存运动神经元1（SMN1）基因的丢失引起的，然而，一个称为SMN2的相同编码基因被保留，但被选择性剪接以产生约90%的截短蛋白。大多数SMA转化和临床前药物开发依赖于使用SMA小鼠来确定SMN蛋白水平的变化。然而，SMA小鼠模型相对严重，SMN诱导化合物的分析受到这些动物早期死亡的混淆。可以在Smn背景下检测SMN蛋白的抗体可以规避这种限制，并允许检查未受影响的杂合动物。在这里，我们描述的单克隆抗SMN抗体，4F11，它特异性地识别人类SMN蛋白的生成和表征。4F11检测SMN 2转基因小鼠和SMA细胞系中的SMN（人）蛋白，但不检测天然Smn（小鼠）蛋白。我们证明了使用4F11检测SMA患者成纤维细胞和健康SMN2转基因小鼠中SMN2衍生的SMN蛋白变化的可行性。因此，该抗体是在患者细胞以及转基因小鼠中检查SMN2诱导疗法的极好工具。
Spinal muscular atrophy (SMA), the leading genetic cause of infant death results from loss of spinal motor neurons causing atrophy of skeletal muscle. SMA is caused by loss of the Survival Motor Neuron 1 (SMN1) gene, however, an identically-coding gene called SMN2 is retained, but is alternatively spliced to produce ~90% truncated protein. Most SMA translational and preclinical drug development has relied on the use of SMA mice to determine changes in SMN protein levels. However, the SMA mouse models are relatively severe and analysis of SMN-inducing compounds is confounded by the early mortality of these animals. An antibody that could detect SMN protein on a Smn background could circumvent this limitation and allow unaffected, heterozygous animals to be examined. Here we describe the generation and characterization of a monoclonal anti-SMN antibody, 4F11, which specifically recognizes human SMN protein. 4F11 detects SMN (human) but not native Smn (mouse) protein in SMN2 transgenic mice and in SMA cell lines. We demonstrate the feasibility of using 4F11 to detect changes in SMN2-derived SMN protein in SMA patient fibroblasts and in healthy SMN2 transgenic mice. This antibody is, therefore, an excellent tool for examining SMN2-inducing therapeutics in patient cells as well as in transgenic mice.
登录
查看更多内容
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：
Monani, UR;Coovert, DD;Burghes, AHM
通讯作者：
Burghes, AHM
影响因子：
30.8
作者：
Lefebvre, S;Burlet, P;Melki, J
通讯作者：
Melki, J
影响因子：
15.9
作者：
Avila, Amy M.;Burnett, Barrington G.;Sumner, Charlotte J.
通讯作者：
Sumner, Charlotte J.
影响因子：
5.7
作者：
Baron-Delage, S;Abadie, A;Beretta, L
通讯作者：
Beretta, L