Development of a new membrane protein expression system to replace the use of Xenopus laevis in novel drug development


Membrane transporter proteins are the primary route for cellular acquisition of essential nutrients, including for malignant cells and pathogens. Therefore, transporters are widely viewed as important current and future targets for the development of novel drugs in the treatment of multiple human diseases. One of the most widespread methods used to research medically relevant transporters has been the South African clawed frog, Xenopus laevis. Use of the frogs involves invasive surgery and removal of the female eggs for studying the physiology and biochemistry of important biomedical transporters. This project proposes to work towards long-term direct replacement of the South African clawed frog in membrane transporter research with an animal-free alternative insect cell Sf21 expression system. The Sf21 system uses cell-extracts to produce membrane proteins for research and drug screening without the use of animal products and is a clonal cell line originally derived from the ovaries of a single Fall Armyworm moth (Spodoptera frugiperda). The Sf21 system will be used to synthesise numerous human transporters of type 2 diabetes mellitus and certain carcinomas.