Transient transfection of mammalian cells using a violet diode laser.
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.
Torres-Mapa
Maria Leilani
ML
University of St. Andrews, Scottish University Physics Alliance (SUPA), School of Physics and Astronomy, North Haugh, St. Andrews, Scotland, United Kingdom. mlt9@st-andrews.ac.uk
Angus
Liselotte
L
Ploschner
Martin
M
Dholakia
Kishan
K
Gunn-Moore
Frank J
FJ
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism* / Q000528:radiation effects*; D006224:Cricetinae; D003412:Cricetulus; D004867:Equipment Design; D019544:Equipment Failure Analysis; D057809:HEK293 Cells; D006801:Humans; D007834:Lasers; D034741:RNA, Small Interfering / Q000235:genetics* / Q000493:pharmacokinetics*; D012666:Semiconductors; D014162:Transfection / Q000295:instrumentation*
D034741:RNA, Small Interfering