Case Studies
SAturated Transposon Analysis in Yeast (SATAY) allows one-step mapping of all genetic loci in which transposons can insert without disrupting essential functions. SATAY is particularly suited to discover loci important for growth under various conditions.
Transposon Screen
SATAY was used to identify synthetic rescue and lethality tdp1-AID wss1Δ genetic interactions, in which the DDI1 gene as one of the strongest negative genetic interactors of the tdp1-AID wss1Δ mutant.
The study discovered several cases of genes that appeared to be essential as a consequence of the specific strain and growth conditions originally used to test the viability of gene deletions.
The transposon insertion mutated 91.5% of nonessential genes and 52.5% of essential genes. The number of transposon insertion sites of most genes remained unchanged following DoxSa treatment at the lower concentration, suggesting that the complexity of the library was maintained during the screening procedure and that technical variabilities had little impact on the result of the screen. The screen revealed that disruption of genes that are required for synthesis of very long-chain fatty acyl-CoA and its presentation to the Cer synthase (Δfat1, Δacb1, Δelo2) rendered the cell resistant to DoxSa. This finding validates that changes in fitness of the mutants in the screen were mainly due to the high toxicity of C26-DoxDHCer.
SATAY (1) reveals positive and negative genetic interactions in single and multiple mutant strains, (2) can identify drug targets, (3) detects not only essential genes, but also essential protein domains, (4) generates both null and other informative alleles.
