Transgenic Insect and Use of Same in Methods for Testing Natural or Synthetic Substances

A method for generating a transgenic insect, comprising:

• subcloning a first exogenous DNA sequence encoding a human membrane transporter protein into an expression vector to obtain a vector comprising the first exogenous DNA sequence;
• introducing the vector comprising the first exogenous DNA sequence into an insect to obtain a stable strain of a transgenic precursor insect;
• and
• crossing the transgenic precursor insect with an insect comprising an expression system matched with the expression vector to obtain a transgenic insect with a functional human membrane transporter protein.

A method for testing a synthetic or natural compound to be studied with respect to the interaction thereof with a human membrane transporter protein, the method comprising:

• selecting a human membrane transporter protein from the group consisting of:
• OCT1, OCT2, OATP1B1, OATP1B3, OAT1, OAT3, MDR1, BSEP, BCRP, MATE1, MATE2, and genetic variants thereof;
• providing at least one embryo of a transgenic insect with the selected human membrane transporter protein;
• introducing a tracer substance into the at least one embryo;
• initially measuring an accumulation of the tracer substance in at least one of 1) cells or tissues of the at least one embryo in which the selected human membrane transporter protein is specifically expressed, and 2) a tissue-specific lumen of the at least one embryo;
• introducing a selected one of a synthetic compound to be studied and a natural compound to be studied into the at least one embryo by at least one of injection into the at least one embryo and incubation of the embryos in a dissolved form of the selected compound;
• subsequently measuring, after introducing the selected compound, an accumulation of the tracer substance in at least one of 1) the cells or tissues of the at least one embryo in which the human membrane transporter protein is specifically expressed, and 2) the tissue-specific lumens of the at least one embryo, with the aid of a selected one of a high-spatial-resolution imaging mass spectrometry method, and a fluorescence microscopy;
• and
• determining, with the aid of the selected one of a high-spatial-resolution imaging mass spectrometry method and fluorescence microscopy, that
• the selected compound interacts with an uptake membrane transporter protein when a reduced accumulation of the tracer substance is measured, in the cells or tissues of the at least one embryo in which the human membrane transporter protein is specifically expressed, in the subsequent measurement compared to the initial measurement,
• the selected compound does not interact with an uptake membrane transporter protein when an unaltered accumulation of the tracer substance is measured, in the cells or tissues of the at least one embryo in which the human membrane transporter protein is specifically expressed, in the subsequent measurement compared to the initial measurement,
• the selected compound interacts with an efflux membrane transporter protein when a reduced accumulation of the tracer substance is measured, in the tissue-specific lumens of the at least one embryo, in the subsequent measurement compared to the initial measurement, and
• the selected compound does not interact with an efflux membrane transporter protein when an unaltered accumulation of the tracer substance is measured, in the tissue-specific lumens of the at least one embryo, in the subsequent measurement compared to the initial measurement.