Ultrasound Mediated Transfection of Yeast for Bio-Sensing Applications

Sonoporation is a facile method of transferring exogenous DNA into cells. In sonoporation, ultrasound is used to create transient pores in cell membranes to allow for the passage of foreign particles. This technique has proven feasible in several cell models, but it does not appear in the literature to have been tested on yeast. Yeast are of particular interest in this study because they are a very robust organism that is ideal for bio-sensing. The ability to easily and cheaply transfect yeast using sonoporation would be a large step towards an effective bio-sensor. Transfection is the process by which DNA external to the cell is passed into the cell and taken up into the nucleus. This process is important because it allows for the development of different types of bio-sensors and detectors. The full mechanism by which sonoporation works is not yet fully elucidated, but it is thought to primarily act through acoustic cavitation. The addition of microbubbles to the solution lowers the cavitation threshold and allows greater transient pore formation in the cell membrane. In this research, the sonoporation of yeast was studied using two cell impermeable fluorescent dyes, FITC-Dextran and Propidium Iodide. Ultrasound was applied to yeast colonies in the presence of microbubbles and fluorescent dyes. Optical and Fluorescence microscopy allows for the characterization of the uptake of FITC-Dextran and Propidium Iodide. The goal of this study is to determine if sonoporation is an efficient and easy way to transfect yeast.