Menolascina Filippo, Fiore Gianfranco, Orabona Emanuele, De Stefano Luca, Ferry Mike, Hasty Jeff, di Bernardo Mario, di Bernardo Diego
TeleThon Institute of Genetics and Medicine (TIGEM), Naples, Italy; Department of Electrical Engineering and Information Technology, University of Naples Federico II, Naples, Italy.
Institute of Microelectronics and Microsystems (IMM), CNR, Naples, Italy.
PLoS Comput Biol. 2014 May 15;10(5):e1003625. doi: 10.1371/journal.pcbi.1003625. eCollection 2014 May.
We describe an innovative experimental and computational approach to control the expression of a protein in a population of yeast cells. We designed a simple control algorithm to automatically regulate the administration of inducer molecules to the cells by comparing the actual protein expression level in the cell population with the desired expression level. We then built an automated platform based on a microfluidic device, a time-lapse microscopy apparatus, and a set of motorized syringes, all controlled by a computer. We tested the platform to force yeast cells to express a desired fixed, or time-varying, amount of a reporter protein over thousands of minutes. The computer automatically switched the type of sugar administered to the cells, its concentration and its duration, according to the control algorithm. Our approach can be used to control expression of any protein, fused to a fluorescent reporter, provided that an external molecule known to (indirectly) affect its promoter activity is available.
我们描述了一种创新的实验和计算方法，用于控制酵母细胞群体中蛋白质的表达。我们设计了一种简单的控制算法，通过将细胞群体中的实际蛋白质表达水平与期望表达水平进行比较，自动调节向细胞中诱导分子的给药。然后，我们基于微流控装置、延时显微镜设备和一组电动注射器构建了一个自动化平台，所有这些都由计算机控制。我们测试了该平台，以迫使酵母细胞在数千分钟内表达所需的固定或随时间变化量的报告蛋白。计算机根据控制算法自动切换给予细胞的糖的类型、浓度和持续时间。只要有一种已知（间接）影响其启动子活性的外部分子可用，我们的方法就可用于控制与荧光报告基因融合的任何蛋白质的表达。
