University of St. Andrews, Scottish University Physics Alliance (SUPA), School of Physics and Astronomy, North Haugh, St. Andrews, Scotland, United Kingdom.
J Biomed Opt. 2010 Jul-Aug;15(4):041506. doi: 10.1117/1.3430730.
We demonstrate the first use of the violet diode laser for transient mammalian cell transfection. In contrast to previous studies, which showed the generation of stable cell lines over a few weeks, we develop a methodology to transiently transfect cells with an efficiency of up to approximately 40%. Chinese hamster ovary (CHO-K1) and human embryonic kidney (HEK293) cells are exposed to a tightly focused 405-nm laser in the presence of plasmid DNA encoding for a mitochondrial targeted red fluorescent protein. We report transfection efficiencies as a function of laser power and exposure time for our system. We also show, for the first time, that a continuous wave laser source can be successfully applied to selective gene silencing experiments using small interfering RNA. This work is a major step towards an inexpensive and portable phototransfection system.
我们展示了紫激光二极管在哺乳动物细胞瞬时转染中的首次应用。与之前需要数周时间才能获得稳定细胞系的研究不同，我们开发了一种方法，可将细胞的瞬时转染效率提高到约 40%。在存在编码靶向线粒体的红色荧光蛋白的质粒 DNA 的情况下，用紧聚焦的 405nm 激光照射中国仓鼠卵巢（CHO-K1）和人胚肾（HEK293）细胞。我们报告了我们的系统中激光功率和照射时间对转染效率的影响。我们还首次表明，连续波激光源可成功应用于使用小干扰 RNA 的选择性基因沉默实验。这项工作是朝着开发廉价、便携的光转染系统迈出的重要一步。
