title: Transient transfection of mammalian cells using a violet diode laser.
abstract: 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.
authors.affiliation: 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
authors.forename: Maria Leilani
authors.initials: ML
authors.lastname: Torres-Mapa
authors.forename: Liselotte
authors.initials: L
authors.lastname: Angus
authors.forename: Martin
authors.initials: M
authors.lastname: Ploschner
authors.forename: Kishan
authors.initials: K
authors.lastname: Dholakia
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Gunn-Moore
mesh_terms: 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*
chemical_list: D034741:RNA, Small Interfering