Genome editing and high-throughput cell analysis platform

基因组编辑和高通量细胞分析平台

• 批准号: RTI-2020-00389
• 负责人: Bechstedt, Susanne
• 金额: $ 10.85万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=693648
• 关键词: Genome; editing; throughput; cell; analysis

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.**

我们的提案要求提供我们迫切需要的设备套件，以满足我们对基因组编辑和高内容成像工作流程的需求。IncuCyte SX1高通量成像平台是一个强大而经济的解决方案，使我们能够使用相衬（无标签）成像和荧光标记探针执行各种活细胞分析。IncuCyte SX1在细胞培养箱中工作，因此保证几乎无干扰，细胞系的长期成像。为了产生IncuCyte SX1检测的敲除、敲除和敲入细胞系，我们需要核因子4D。该仪器使用电脉冲将DNA， RNA和蛋白质直接输送到细胞质和细胞核中，同时保持高细胞活力。该技术是为CRISPR基因组编辑提供纯化的Cas9核酸酶与sgRNA双工的基准。成功的基因组编辑需要高达99%的转染效率，并结合大多数细胞系的高活力。在细胞核感染的下游，通过稀释克隆选择单细胞克隆，将使用IncuCyte SX1进行监测。最后，我们需要一个CellDrop消耗性免费细胞计数器，作为一个实用和经济的解决方案，以跟上我们HQPs处理的细胞培养量的增加。******所要求的设备将使我们的实验室能够在竞争激烈的国际研究环境中继续取得成功，并产生高影响力的工作。******这些设备将由我们的hqp使用，确保有机会对高度先进的技术进行实践培训。独特的设备和经过深思熟虑的技术知识将使我们的hqp在他们的项目以及未来的职业选择中具有很强的竞争力。***设备将有利于HQPs从我们的实验室和其他实验室在部门以及蒙特利尔研究界