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An Effort to Use Human-Based Exome Capture Methods to Analyze Chimpanzee and Macaque Exomes

机译:使用基于人的外显子组捕获方法来分析黑猩猩和猕猴外显子组的努力

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摘要

Non-human primates have emerged as an important resource for the study of human disease and evolution. The characterization of genomic variation between and within non-human primate species could advance the development of genetically defined non-human primate disease models. However, non-human primate specific reagents that would expedite such research, such as exon-capture tools, are lacking. We evaluated the efficiency of using a human exome capture design for the selective enrichment of exonic regions of non-human primates. We compared the exon sequence recovery in nine chimpanzees, two crab-eating macaques and eight Japanese macaques. Over 91% of the target regions were captured in the non-human primate samples, although the specificity of the capture decreased as evolutionary divergence from humans increased. Both intra-specific and inter-specific DNA variants were identified; Sanger-based resequencing validated 85.4% of 41 randomly selected SNPs. Among the short indels identified, a majority (54.6%–77.3%) of the variants resulted in a change of 3 base pairs, consistent with expectations for a selection against frame shift mutations. Taken together, these findings indicate that use of a human design exon-capture array can provide efficient enrichment of non-human primate gene regions. Accordingly, use of the human exon-capture methods provides an attractive, cost-effective approach for the comparative analysis of non-human primate genomes, including gene-based DNA variant discovery.
机译:非人类灵长类动物已经成为研究人类疾病和进化的重要资源。非人类灵长类物种之间和之内的基因组变异的表征可以促进遗传学定义的非人类灵长类动物疾病模型的发展。但是,缺少能够加快这类研究的非人类灵长类动物特异性试剂,例如外显子捕获工具。我们评估了使用人类外显子组捕获设计对非人类灵长类动物的外显子区域进行选择性富集的效率。我们比较了九只黑猩猩,两只食蟹的猕猴和八只日本猕猴的外显子序列恢复情况。超过91%的目标区域是在非人类灵长类动物样品中捕获的,尽管捕获的特异性随着与人类进化差异的增加而降低。鉴定出种内和种间DNA变体。基于Sanger的重新测序验证了41个随机选择的SNP中的85.4%。在鉴定出的短插入缺失中,大多数变异(54.6%–77.3%)导致3个碱基对的变化,这与针对移码突变的选择预期相符。综上所述,这些发现表明使用人类设计的外显子捕获阵列可以提供非人类灵长类动物基因区域的有效富集。因此,使用人类外显子捕获方法为非人类灵长类动物基因组的比较分析(包括基于基因的DNA变体发现)提供了一种有吸引力的,具有成本效益的方法。

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