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 -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.
Bibliographical noteFunding Information:
We thank Dr. David Stevenson for useful discussions. We thank the United Kingdom Engineering and Physical Sciences Research Council and Biotechnology and Biological Sciences Research Council for funding. MLT acknowledges the support of a SUPA Prize Studentship. KD is a Royal Society-Wolfson Merit Award Holder. FGM and KD contributed equally to this work.
All Science Journal Classification (ASJC) codes
- Electronic, Optical and Magnetic Materials
- Atomic and Molecular Physics, and Optics
- Biomedical Engineering