High performance nucleofection unit for difficult to manipulate cell types

我们对动物或人类发生的事情的了解，大部分来自对这些复杂生物体中的单个成分或块的研究。就像汽车修理工拆卸发动机的所有部件一样，生物学家也会拆卸和更换活体的各个部件，以了解每个部件的作用。细胞是所有已知生物体的基本结构、功能和生物单位。为了研究细胞如何对环境做出反应，以及它们如何相互交流，必须删除，修改或添加组件。众所周知，并非所有的通信事件和/或相互作用都发生在所有类型的细胞中。因此，为了充分了解某些细胞过程的功能，必须研究感兴趣的细胞类型，其中一些非常难以操作。因此，本申请的目标是整合一种先进的技术，该技术允许将外源物质（cDNA、siRNA和miRNA）容易地引入这些难以操作的细胞类型中。核转染是一种通过电脉冲将材料引入难以转染的细胞类型的技术。虽然这个概念并不是新技术（实际上是在1965年发现的），但Amaxa 4D Nucleofector是一个重大进步，它允许操纵许多细胞类型，同时保持它们的存活。它是将物质（核酸）引入非分裂细胞（如神经元和静息血细胞）的有效手段。这种方法是对需要病毒的其他技术的改进，这些技术昂贵，具有安全风险并且通常缺乏可靠性。 Vande Velde、Leclerc、Arabel和Fernandes的团队专注于了解大脑和血液不同细胞类型中的特定通信系统。这些NSERC资助（或更新）的研究项目如果不能操纵相关细胞类型中的相应细胞过程，就无法取得进展。所要求的设备对于高质量的HQP的持续培训和推进我们对基本生物过程的集体理解至关重要。

Much of what we know about what is happening in animals or humans derives from studies investigating individual components or blocks included in these complex organisms. Like a car mechanic dismantling all pieces of an engine, biologists dismantle and change pieces of living bodies to enlighten the role of each component. Cells are the basic structural, functional and biological unit of all known living organisms. To study how cells react to their environment and how they communicate between each other it is essential to remove, modify or add components to them. It is well recognized that not all communication events and/or interactions occur in all types of cells. Thus, in order to fully understand how certain cellular processes function, one must study in the cell type of interest, some of which are extremely difficult to manipulate. Hence, the goal of this application is to integrate an advanced technology that permits facile introduction of exogenous material (cDNAs, siRNAs and miRNAs) into these difficult-to-manipulate cell types. Nucleofection is a technique that introduces material into difficult-to-transfect cell types via an electrical pulse. While the concept is not new technology (actually discovered in 1965), the requested Amaxa 4D Nucleofector, is a major advance that permits the manipulation of numerous cell types while keeping them alive. It is an efficient means to introduce material (nucleic acids) into non-dividing cells such as neurons and resting blood cells. This methodology is an improvement over other techniques requiring viruses, which are expensive, have safety risks and often lack reliability. The teams of Vande Velde, Leclerc, Arbour and Fernandes are focused on understanding specific communication systems in different cell types of the brain and the blood. These NSERC-funded (or to be renewed) research programs cannot advance without the ability to manipulate the respective cellular processes in the relevant cell types. The requested equipment is essential for the continued training of high quality HQP and to advancing our collective understanding of fundamental biological processes.