Genetic Screening using High-Throughput Barcode Sequencing of Saccharomyces cerevisiae Knockout Collection: Search for Regulators of mtDNA Intracellular Selection

Abstract

We searched for genes whose deletion affects mitochondrial heteroplasmy in the yeast Saccharomyces cerevisiae. In order to do this, we crossed a yeast knockout collection pool (rho⁺) with a rho^– strain containing a large deletion in its mtDNA. The resulting mixture of cells was plated onto selective media that made it possible to distinguish between diploid cells that had lost and retained wild-type mtDNA as a result of such crossing. From the resulting pools (initial one, cells that lose rho⁺ mtDNA, and those that retained it), we isolated DNA and determined the proportion of deletion strains using high-throughput sequencing. This made it possible to identify functional groups of genes that reduce the ability of yeast cells to maintain rho⁺ mtDNA in a state of heteroplasmy with rho^– mtDNA.