Wellcome Trust Sanger Institute, Hinxton, UK.
European Molecular Biology Laboratory, European Bioinformatics Institute, Wellcome Genome Campus, Hinxton, UK.
Nat Genet. 2018 Jan;50(1):54-61. doi: 10.1038/s41588-017-0005-8. Epub 2017 Dec 11.
Induced pluripotent stem cells (iPSCs), and cells derived from them, have become key tools for modeling biological processes, particularly in cell types that are difficult to obtain from living donors. Here we present a map of regulatory variants in iPSC-derived neurons, based on 123 differentiations of iPSCs to a sensory neuronal fate. Gene expression was more variable across cultures than in primary dorsal root ganglion, particularly for genes related to nervous system development. Using single-cell RNA-sequencing, we found that the number of neuronal versus contaminating cells was influenced by iPSC culture conditions before differentiation. Despite high differentiation-induced variability, our allele-specific method detected thousands of quantitative trait loci (QTLs) that influenced gene expression, chromatin accessibility, and RNA splicing. On the basis of these detected QTLs, we estimate that recall-by-genotype studies that use iPSC-derived cells will require cells from at least 20-80 individuals to detect the effects of regulatory variants with moderately large effect sizes.
诱导多能干细胞(iPSCs)及其衍生细胞已成为模拟生物过程的重要工具,特别是在难以从活体供体中获得的细胞类型中。在这里,我们基于 123 种 iPSC 向感觉神经元命运的分化,提供了 iPSC 衍生神经元中调节变体的图谱。与原代背根神经节相比,基因表达在培养物之间的变异性更大,特别是与神经系统发育相关的基因。使用单细胞 RNA 测序,我们发现单细胞与污染细胞的数量受到分化前 iPSC 培养条件的影响。尽管分化诱导的变异性很高,但我们的等位基因特异性方法检测到了数千个影响基因表达、染色质可及性和 RNA 剪接的数量性状基因座(QTL)。基于这些检测到的 QTL,我们估计使用 iPSC 衍生细胞的基于基因型召回的研究将需要至少 20-80 个人的细胞来检测具有中等效应大小的调节变体的影响。