Simultaneous quantification of mRNA and protein in single cells reveals post-transcriptional effects of genetic variation.
Simultaneous quantification of mRNA and protein in single cells reveals post-transcriptional effects of genetic variation.
复制标题
单细胞中mRNA和蛋白质的同时定量揭示了遗传变异的转录后效应。
DOI：
10.7554/elife.60645
复制
发表时间：
2020-11-16
期刊：
影响因子：
7.7
通讯作者：
Albert FW
中科院分区：
文献类型：
--
作者：
Brion C;Lutz SM;Albert FW
反式作用的DNA变异体可能会特异性地影响整个基因组中基因的mRNA或蛋白质水平。然而，之前的工作比较了在不同研究中绘制的反式作用基因座，其中许多研究的统计能力有限。在这里，我们开发了一个基于CRISPR的系统，通过双荧光报告器在酿酒酵母的单个活细胞中同时定量给定基因的mRNA和蛋白质。在来自两个遗传差异菌株杂交的重组细胞的大群体中，我们定位了影响10个基因表达的86个反式作用位点。只有不到20%的这些基因座对同一基因的mRNA和蛋白质产生了一致的影响。大多数基因座影响的是蛋白质，而不是特定基因的mRNA.其中一个基因座含有YAK1激酶基因的提前终止变异，该变异对数十个基因的蛋白质或mRNA有特定影响。这些结果显示了复杂的转录后遗传效应对基因表达的影响。
Trans-acting DNA variants may specifically affect mRNA or protein levels of genes located throughout the genome. However, prior work compared trans-acting loci mapped in separate studies, many of which had limited statistical power. Here, we developed a CRISPR-based system for simultaneous quantification of mRNA and protein of a given gene via dual fluorescent reporters in single, live cells of the yeast Saccharomyces cerevisiae. In large populations of recombinant cells from a cross between two genetically divergent strains, we mapped 86 trans-acting loci affecting the expression of ten genes. Less than 20% of these loci had concordant effects on mRNA and protein of the same gene. Most loci influenced protein but not mRNA of a given gene. One locus harbored a premature stop variant in the YAK1 kinase gene that had specific effects on protein or mRNA of dozens of genes. These results demonstrate complex, post-transcriptional genetic effects on gene expression.