The Jackson Laboratory, Bar Harbor, ME 04609, USA.
Hum Mol Genet. 2012 Oct 15;21(20):4431-47. doi: 10.1093/hmg/dds285. Epub 2012 Jul 16.
A number of mouse models for spinal muscular atrophy (SMA) have been genetically engineered to recapitulate the severity of human SMA by using a targeted null mutation at the mouse Smn1 locus coupled with the transgenic addition of varying copy numbers of human SMN2 genes. Although this approach has been useful in modeling severe SMA and very mild SMA, a mouse model of the intermediate form of the disease would provide an additional research tool amenable for drug discovery. In addition, many of the previously engineered SMA strains are multi-allelic by design, containing a combination of transgenes and targeted mutations in the homozygous state, making further genetic manipulation difficult. A new genetic engineering approach was developed whereby variable numbers of SMN2 sequences were incorporated directly into the murine Smn1 locus. Using combinations of these alleles, we generated an allelic series of SMA mouse strains harboring no, one, two, three, four, five, six or eight copies of SMN2. We report here the characterization of SMA mutants in this series that displayed a range in disease severity from embryonic lethal to viable with mild neuromuscular deficits.
许多脊髓性肌萎缩症(SMA)的小鼠模型是通过在 Smn1 基因座上进行靶向缺失突变,同时转染不同拷贝数的人 SMN2 基因来重现人类 SMA 的严重程度而遗传工程构建的。尽管这种方法在模拟严重 SMA 和非常轻度 SMA 方面很有用,但疾病中间形式的小鼠模型将提供另一种可用于药物发现的研究工具。此外,许多以前设计的 SMA 品系是多等位基因的,在纯合状态下包含转基因和靶向突变的组合,使得进一步的遗传操作变得困难。开发了一种新的遗传工程方法,其中将可变数量的 SMN2 序列直接整合到鼠 Smn1 基因座中。使用这些等位基因的组合,我们生成了一系列携带零、一、二、三、四、五、六或八个 SMN2 拷贝的 SMA 小鼠品系。我们在这里报告了该系列中 SMA 突变体的特征,这些突变体的疾病严重程度从胚胎致死到存活但有轻微的神经肌肉缺陷不等。