High efficiency DNA transfer into mammalian and non-mammalian cells

将 DNA 高效转移至哺乳动物和非哺乳动物细胞中

• 批准号: 375276-2009
• 负责人: Damjanovski, Sashko
• 金额: $ 2.84万
• 依托单位: University of Western Ontario
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2008
• 资助国家: 加拿大
• 起止时间: 2008-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=412123
• 关键词: efficiency; DNA; transfer; into; mammalian

In addition to using whole animals, such as frogs, mice or fish, to answer biological questions, there are numerous cells from such animals that can be used on their own to ask different questions that might be possible with a whole animal. Many such in vitro cell lines exist and they are invaluable research tools as the variety of cell lines allows them to be used in complementary approaches to examine different cellular functions (migration, differentiation, signalling, transformation). The advent of genomic sequencing, CHIP screening, and other technologies, coupled with advances with imaging systems, has now fostered in an era of science, where there are opportunities to investigate a protein's function(s) as never before. However, despite advances, many useful cell lines have distinct and often quite low DNA transfer or transfection efficiencies. The ability to introduce exogenous DNA or RNA into a cell is pivotal to address many of the most pertinent questions as to how specific proteins function, and different cell lines have different abilities to take up exogenous DNA. This is problematic when the functions of proteins are compared between different cell lines. Indeed optimized transfection protocols that do exist for a given cell line are often unlike, and introduce variability when several cell lines are used and compared. As such, a high efficiency DNA transfer system that can be used for both mammalian and non-mammalian cell transfections would allow for standardization between cell lines, and would thus facilitate an integration of data.

除了使用整个动物，如青蛙、老鼠或鱼来回答生物学问题外，还有大量来自这些动物的细胞可以单独用来提出可能对整个动物都可能提出的不同问题。存在许多这样的体外细胞系，它们是无价的研究工具，因为细胞系的多样性使它们能够用于互补的方法来检查不同的细胞功能(迁移、分化、信号、转化)。基因组测序、芯片筛选和其他技术的出现，加上成像系统的进步，现在促进了一个科学时代的到来，在这个时代，有机会以前所未有的方式研究蛋白质的功能(S)。然而，尽管取得了进展，许多有用的细胞系具有明显的DNA转移或转染率，而且往往相当低。将外源DNA或RNA导入细胞的能力对于解决许多最相关的问题至关重要，这些问题涉及特定蛋白质的功能，不同的细胞系有不同的能力来吸收外源DNA。当蛋白质的功能在不同细胞系之间进行比较时，这是有问题的。事实上，对于给定的细胞系确实存在的优化的转染法通常是不同的，并且在使用和比较几个细胞系时引入了可变性。因此，可以用于哺乳动物和非哺乳动物细胞转基因的高效DNA转移系统将允许细胞系之间的标准化，从而将促进数据的整合。