PT  - JOURNAL ARTICLE
AU  - Christian Brion
AU  - Sheila Lutz
AU  - Frank W. Albert
TI  - Simultaneous quantification of mRNA and protein in single cells reveals post-transcriptional effects of genetic variation
AID  - 10.1101/2020.07.02.185413
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.07.02.185413
4099  - http://biorxiv.org/content/early/2020/07/03/2020.07.02.185413.short
4100  - http://biorxiv.org/content/early/2020/07/03/2020.07.02.185413.full
AB  - 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.