Genetics of stroke vulnerability in C57BL/6 mouse substrains

中风仍然是一种重要的人类疾病，理解遗传因素 促进其对特定个人的影响最终将是针对性干预措施的关键。 在人群中进行的研究通常涉及影响中风发生风险的因素。 在实验模型和平行映射中很容易检验对中风严重性的影响 在老鼠和人类群体中的易感基因座的研究支持了这种 分析。我们最近发现中风易感性在密切的人群中存在明显的差异 C57BL/6小鼠的相关亚系。在J和ByJ亚系中可以看到较小的梗塞， 后者起源于早期的N世系。相比之下，更大的梗塞发生在 随后获得了NCrSLc、NCr1和Nj亚株。初步结果显示， 较大的梗塞表型是遗传的显性特征，表明是单一原因突变。 这些近乎同基因的亚株群体只包含有限的分离基因库。 可能导致中风严重程度差异的变异。 我们建议在C57BL/6中确定调节卒中易感性的基因变异 亚株。AIM 1将为ByJ、NCrslc和NCr1生成全面的序列数据 亚株，目前还没有这样的资源。因为BYJ和NCRSLC是最 密切相关的亚株在中风易感性上有所不同，这将定义 候选变种。AIM 2将使用高覆盖率的基因组序列和密集的标记面板 已可用于J和NJ亚株以倒数形式绘制影响梗死体积的位置 JxNJ和NJxJ F2杂交后代群体遭受永久性局灶性缺血。 该基因座将利用ByJ和NCr1亚株的F2杂交得到确认。目标3，使用CRISPR-- 修复每个候选变异的Cas9方法，将建立单一的遗传因素，即 C57BL/6亚型间卒中易感性变异的主要来源。

Stroke remains a significant human disease, and understanding the genetic factors that contribute to its impact on specific individuals will ultimately be critical in targeting interventions. Studies in human populations generally address factors affecting the risk of stroke occurrence. Impacts on stroke severity are readily examined in experimental models, and parallel mapping of vulnerability loci in mouse and human populations support the translational potential of such analyses. We have recently identified marked divergence of stroke vulnerability among closely related substrains of the C57BL/6 mouse. Smaller infarcts are seen in the J and ByJ substrains, the latter of which originated early in the N lineage. In contrast, larger infarcts are observed in subsequently derived NCrSlc, NCrl and NJ substrains. Preliminary results establish that the larger infarct phenotype is inherited as a dominant trait, indicating a single causal mutation. These nearly coisogenic substrain populations contain only a limited pool of segregating genetic variants that could underlie the difference in stroke severity. We propose to identify the gene variant that modulates stroke vulnerability among C57BL/6 substrains. Aim 1 will generate comprehensive sequence data for ByJ, NCrSlc and NCrl substrains, for which such resources are not yet available. Since ByJ and NCrSlc are the most closely related substrains to differ in stroke vulnerability, this will define the minimum list of candidate variants. Aim 2 will use high-coverage genomic sequences and a dense marker panel already available for J and NJ substrains to map the locus impacting infarct volume in reciprocal populations of JxNJ and NJxJ F2 intercross progeny subjected to permanent focal ischemia. This locus will be confirmed using F2 crosses of ByJ and NCrl substrains. Aim 3, using CRISPR- Cas9 methodology to repair each candidate variant, will establish the single genetic factor that is the main source of variation in stroke vulnerability among C57BL/6 substrains.