Genome editing and high-throughput cell analysis platform

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Our proposal requests the equipment suite we urgently need to keep up with our demand for genome editing and high-content imaging workflows. The IncuCyte SX1 high-throughput imaging platform is a powerful yet economical solution that allows us to perform a wide variety of live cell assays using phase contrast (label free) imaging and fluorescently labeled probes. The IncuCyte SX1 works inside a cell culture incubator and therefore guarantees virtually disturbance free, long-term imaging of cell lines. To produce the knock-out, knock-down, and knock-in cell lines to be assayed by the IncuCyte SX1, we request the Nucleofector 4D. The instrument uses electrical pulses to deliver DNA, RNA, and protein directly into the cytosol and nucleus while maintaining high cell viability. The technology is the benchmark in delivering purified Cas9 nuclease duplexed with sgRNA for CRISPR genome editing. Up to 99% transfection efficiency combined with high viability for most cell lines is required for successful genome editing. Downstream of nucleofection a selection of single cell clones by dilution cloning will be monitored using the IncuCyte SX1. Lastly, we are requesting a CellDrop consumable free cell counter as a practical and economical solution to keep up with the increased cell culture volume handled by our HQPs.******The equipment requested will enable our labs to continue to succeed in a highly competitive international research environment and produce high impact work. ******The equipment will be used by our HQPs, ensuring hands-on training opportunities on highly advanced technologies. The unique equipment and thought-after technology knowledge will enable our HQPs to be very competitive in their projects as well as their future career choices.***The equipment will benefit HQPs from our labs and from other labs in the department as well as the Montreal research community.**