Yeast double transporter-gene deletion library for identification of xenobiotic carriers

The routes of uptake and efflux should be considered when developing new drugs so that they can effectively address their intracellular targets. In general rule, drugs appear to enter cells via protein carriers that normally carry nutrients or metabolites. A previously developed pipeline that searched for drug transporters using Saccharomyces cerevisiae mutants carrying single-gene deletions identified import routes for most compounds tested. However, due to the redundancy of transporter functions, we proposed that this methodology could be improved by utilizing double-mutant strains in both low- and high-throughput screens. We constructed a library of over 14,000 strains harboring double-deletions of 122 non-essential genes encoding plasma membrane transporters and performed high-throughput screens to identify possible drug import and efflux routes for 23 compounds.

在研发新型药物时，需考量药物的摄取与外排通路，以确保其能有效作用于细胞内靶点。一般而言，药物通常通过负责转运营养物质或代谢物的蛋白质载体进入细胞。此前一项利用携带单基因缺失的酿酒酵母（Saccharomyces cerevisiae）突变体筛选药物转运蛋白的研究流程，已为多数受试化合物鉴定出摄取通路。然而，由于转运蛋白功能存在冗余性，我们提出可通过在低通量与高通量筛选中使用双突变菌株，来优化该研究方法。我们构建了包含14000余株菌株的文库，这些菌株均携带有122个编码质膜转运蛋白的非必需基因的双缺失突变，并通过高通量筛选，为23种受试化合物鉴定了潜在的药物摄取与外排通路。