Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health Bethesda, MD, USA.
Front Cell Dev Biol. 2015 Jan 9;2:74. doi: 10.3389/fcell.2014.00074. eCollection 2014.
Non viral-mediated transfection of plasmid DNA provides a fast and reliable way to express various transgenes in selected cell populations in live animals. Here, we show an improvement of a previously published method that is based on injecting plasmid DNA into the ductal system of the salivary glands in live rats. Specifically, using complexes between plasmid DNA and polyethyleneimine (PEI) we show that the expression of the transgenes is directed selectively to the salivary acinar cells. PEI does not affect the ability of cells to undergo regulated exocytosis, which was one of the main drawbacks of the previous methods. Moreover PEI does not affect the proper localization and targeting of transfected proteins, as shown for the apical plasma membrane water channel aquaporin 5 (AQP5). Overall, this approach, coupled with the use of intravital microscopy, permits to conduct localization and functional studies under physiological conditions, in a rapid, reliable, and affordable fashion.
非病毒介导的质粒 DNA 转染为在活体动物中选择的细胞群体中表达各种转基因提供了一种快速可靠的方法。在这里,我们展示了一种基于将质粒 DNA 注射到活体大鼠唾液腺导管系统的先前发表方法的改进。具体来说,我们使用质粒 DNA 和聚乙烯亚胺 (PEI) 之间的复合物表明,转基因的表达被选择性地引导至唾液腺泡细胞。PEI 不会影响细胞进行调节性胞吐的能力,这是先前方法的主要缺点之一。此外,PEI 不会影响转染蛋白的适当定位和靶向,如顶端质膜水通道 aquaporin 5 (AQP5) 所示。总的来说,这种方法与活体显微镜的使用相结合,允许以快速、可靠和经济实惠的方式在生理条件下进行定位和功能研究。