Yeast Double Transporter Gene Deletion Library for Identification of Xenobiotic Carriers in Low or High Throughput

Abstract

Our library of double transporter deletion strains is a powerful tool for rapid identification of potential drug import and export routes, which can aid in determining the chemical groups necessary for transport via specific carriers. This information may be translated into a better design of drugs for optimal absorption by target tissues and the development of drugs whose utility is less likely to be compromised by the selection of resistant mutants.

Analysis of a Library of Escherichia coli Transporter Knockout Strains to Identify Transport Pathways of Antibiotics

“…Further validation of predicted effects can be achieved by use of the ‘ASKA’ overexpression collection [ 34 ], where, if the opposite effect is observed (e.g., resistance in an overexpression strain and sensitivity in a knockout strain), this could be seen as providing further evidence that the compound is indeed a substrate of the given transporter [ 19 , 35 , 36 ]. Given the large degree of redundancy in transporters (i.e., a given substrates may be transported by several transporters), more robust follow-up results could be achieved through investigation of double knockouts, as has recently been demonstrated in yeast [ 37 ].…”

A Chemogenomic Toolkit to Evaluate the “Ins and Outs” of Yeast Plasma Membrane Transporters

“…Given these complexities with membrane transporters, Almeida and colleagues ( 9 ) constructed a double-deletion library of 122 nonessential transmembrane transporters in all possible combinations. The double-deletion strains were obtained by carrying out crosses between strains harboring single transporter gene deletions, each carrying a specific replacement cassette, kan MX or nat MX.…”