期刊
JOURNAL OF BIOMEDICAL OPTICS
卷 15, 期 4, 页码 -出版社
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.3430730
关键词
photoporation; phototransfection; gene transfection; gene knockdown; violet diode
资金
- United Kingdom Engineering and Physical Sciences Research Council
- Biotechnology and Biological Sciences Research Council
- SUPA
- Royal Society
- Engineering and Physical Sciences Research Council [EP/D04877X/1] Funding Source: researchfish
- EPSRC [EP/D04877X/1] Funding Source: UKRI
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 similar to 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. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3430730]
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