Department of Genetic Medicine and Development, University of Geneva Medical School, Geneva, 1211 Switzerland.
Nature. 2010 Apr 1;464(7289):773-7. doi: 10.1038/nature08903. Epub 2010 Mar 10.
Gene expression is an important phenotype that informs about genetic and environmental effects on cellular state. Many studies have previously identified genetic variants for gene expression phenotypes using custom and commercially available microarrays. Second generation sequencing technologies are now providing unprecedented access to the fine structure of the transcriptome. We have sequenced the mRNA fraction of the transcriptome in 60 extended HapMap individuals of European descent and have combined these data with genetic variants from the HapMap3 project. We have quantified exon abundance based on read depth and have also developed methods to quantify whole transcript abundance. We have found that approximately 10 million reads of sequencing can provide access to the same dynamic range as arrays with better quantification of alternative and highly abundant transcripts. Correlation with SNPs (small nucleotide polymorphisms) leads to a larger discovery of eQTLs (expression quantitative trait loci) than with arrays. We also detect a substantial number of variants that influence the structure of mature transcripts indicating variants responsible for alternative splicing. Finally, measures of allele-specific expression allowed the identification of rare eQTLs and allelic differences in transcript structure. This analysis shows that high throughput sequencing technologies reveal new properties of genetic effects on the transcriptome and allow the exploration of genetic effects in cellular processes.
基因表达是一个重要的表型,它可以反映遗传和环境对细胞状态的影响。以前,许多研究都使用定制和商业可用的微阵列来识别基因表达表型的遗传变异。第二代测序技术现在为转录组的精细结构提供了前所未有的访问途径。我们对 60 名欧洲血统的扩展 HapMap 个体的转录组 mRNA 部分进行了测序,并将这些数据与 HapMap3 项目中的遗传变异结合起来。我们根据读取深度量化了外显子丰度,并且还开发了量化整个转录本丰度的方法。我们发现,大约 1000 万条测序读取可以提供与具有更好的替代和高度丰富转录本定量的阵列相同的动态范围。与阵列相比,与 SNPs(单核苷酸多态性)的相关性导致 eQTLs(表达数量性状基因座)的发现更大。我们还检测到大量影响成熟转录本结构的变体,表明负责可变剪接的变体。最后,等位基因特异性表达的测量允许鉴定罕见的 eQTLs 和转录本结构中的等位基因差异。这项分析表明,高通量测序技术揭示了遗传对转录组的影响的新特性,并允许探索细胞过程中的遗传效应。