Photothermally activated protein-producing liposomes using gold nanorods

Abstract

Therapeutic proteins are used for various kinds of treatment. However, treatment of therapeutic proteins has limitation which is un-wanted side effects. To overcome the limitation, controlling expression of proteins is essential. Cell-free protein synthesis (CFPS) is the technology for expressing proteins not inside cells. Various kinds of components for protein expression are prepared and mixed with DNA, for example, amino acids, ions, kinases, and cell extracts. By using this synthesis system, expressing proteins artificially without cell can be possible. However, controlling expression of proteins is hard to achieve because DNA undergoes transcription immediately when they mixed with expression components. RNA thermometer (RNAt), thermo-sensitive riboswitch, is the technology for controlling protein expression. Unless RNAt reaches to proper temperature, they don’t undergo translation process. Here, we use RNAt for controlling protein translation. To activate translation of RNAt, we use photo-thermal effect by using gold nanorods (GNRs) with NIR laser. GNRs were photo-excited by NIR laser, which can penetrate the tissue well, therefore we can achieve translation of RNAt when we stimulate. To deliver and express proteins at target site, we loaded cell-free protein synthesis system into liposome which is safe delivery system for cargos. Finally, we confirmed protein translation of RNAt inside liposome using GNRs by photo-thermal effects with NIR laser.