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bioRxiv · 10.1101/2020.07.02.185413

Simultaneous quantification of mRNA and protein in single cells reveals post-transcriptional effects of genetic variation

Brion, C.·Lutz, S.·Albert, F. W.
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Abstract

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 studies performed separately or with 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 such 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.One sentence summary A CRISPR-based dual reporter assay enables genetic mapping of DNA variants that specifically affect mRNA or protein levels in trans.Competing Interest StatementThe authors have declared no competing interest.View Full Text

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Brion, C., Lutz, S., Albert, F. W.. 2020-07-03. Simultaneous quantification of mRNA and protein in single cells reveals post-transcriptional effects of genetic variation. https://doi.org/10.1101/2020.07.02.185413

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