Cell-based and biochemical high-throughput screening in 1536-well plates
Speaker: Dr. Gene Ananiev (University of Wisconsin, Dr. Carl Peters (BMG LABTECH), Dr. Iain Russell (Labycte)
In our latest webinar, speaker Dr. Gene Ananiev from the Small Molecule Screening Facility at the University of Wisconsin Madison, discusses two examples of high-throughput screening in 1536-well plates, presenting methodology for assay miniaturisation and optimisation.
The two projects were set up for customers and include a cell-based luciferase reporter assay and an AlphaScreen-based screen for protein-protein interactions.
In the first project, CRISPR/Cas9 was used to knock in a Nano luciferase (Nluc) gene into the endogenous human FMR1 gene in neural progenitor cells. This was done in order to create a robust FMR1-Nluc reporter hiPS cell line. Functional assays confirmed that the Nluc activities accurately report FMR1 gene expression levels. Moreover, neural progenitor cells derived from this cell line were optimised for high-throughput screening.
In the second project, the focus was on antibiotic-resistant bacterial infections. As antibiotic resistance is on the rise worldwide, there is an urgent need to identify novel classes of antibiotics that can overcome resistance mechanisms. In this study, nine efficient inhibitors have been identified. They produced response curves with IC50 values of <40 μM. Additionally, two compounds directly bound to PriA, demonstrating the success of this screening strategy. Using a miniaturised AlphaScreen test, costs were drastically reduced and throughput for protein-protein interaction was increased.
In addition to Dr. Ananiev, Dr. Carl Peters from BMG LABTECH and Dr. Iain Russell from Labcyte highlight the microplate reader and dispensing solution that have been used in the projects.