High Throughput Cell Assays
Investigators: Laura Gonzalez, Ph.D. and Michael Gaskin
Collaborators: Holly Yin, Ph.D. (T-Gen)
The ability to manipulate the expression of proteins within cells (either increasing it by protein overexpression or decreasing it by using RNA interference (RNAi)) provides a powerful tool to understand the function of proteins in mammalian cells. With over 22,000 proteins represented in the human genome, studies of all (or a subset) of these proteins requires robotic assistance to handle the cells, screening, and data quantification. Furthermore, a robotic system maintains the quality and consistency between experiments required to produce meaningful results.
Our goal was to develop a high throughput, cell-based, robotic screening pipeline using the RNAi Collection (TRC), a collection of >150,000 short hairpin RNAs (shRNAs) that can individually eliminate the expression of a particular gene within cells, and other overexpression plasmids created in our center and others. Through Thermo Scientific, we acquired and upgraded the current integrated robotic equipment to develop a fully automated platform for the large scale screening of many genes both by increased expression through cDNAs and decreased expression through shRNAs. Images of the completed automation set up in Figure 1.Based at T-Gen, the system provides screening services to both TGen and the ASU community. and is currently being used in our center to study p53 and its co-drivers in breast cancer.