title: Delivery of proteins to mammalian cells via gold nanoparticle mediated laser transfection.
abstract: Nanoparticle laser interactions are in widespread use in cell manipulation. In particular, molecular medicine needs techniques for the directed delivery of molecules into mammalian cells. Proteins are the final mediator of most cellular cascades. However, despite several methodical approaches, the efficient delivery of proteins to cells remains challenging. This paper presents a new protein transfection technique via laser scanning of cells previously incubated with gold nanoparticles. The laser-induced plasmonic effects on the gold nanoparticles cause a transient permeabilization of the cellular membrane, allowing proteins to enter the cell. Applying this technique, it was possible to deliver green fluorescent protein into mammalian cells with an efficiency of 43%, maintaining a high level of cell viability. Furthermore, a functional delivery of Caspase 3, an apoptosis mediating protein, was demonstrated and evaluated in several cellular assays. Compared to conventional protein transfection techniques such as microinjection, the methodical approach presented here enables high-throughput transfection of about 10 000 cells per second. Moreover, a well-defined point in time of delivery is guaranteed by gold nanoparticle mediated laser transfection, allowing the detailed temporal analysis of cellular pathways and protein trafficking.
authors.affiliation: Biomedical Optics Department, Laser Zentrum Hannover e.V., Hollerithallee 8, 30419 Hannover, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Heinemann
authors.forename: S
authors.initials: S
authors.lastname: Kalies
authors.forename: M
authors.initials: M
authors.lastname: Schomaker
authors.forename: W
authors.initials: W
authors.lastname: Ertmer
authors.forename: H
authors.initials: H
authors.lastname: Murua Escobar
authors.forename: H
authors.initials: H
authors.lastname: Meyer
authors.forename: T
authors.initials: T
authors.lastname: Ripken
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism* / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003584:Cytological Techniques; D004285:Dogs; D006046:Gold / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism* / Q000494:pharmacology; D007834:Lasers; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000633:toxicity
chemical_list: D049452:Green Fluorescent Proteins; D006046:Gold; D053148:Caspase 3