Co-transfection of plasmid DNA and laser-generated gold nanoparticles does not disturb the bioactivity of GFP-HMGB1 fusion protein
Creators: name: Petersen, Svea
name: Soller, Jan T.
name: Wagner, Siegfried
name: Richter, Andreas
name: Bullerdiek, Jörn
name: Nolte, Ingo
name: Barcikowski, Stephan
name: Murua Escobar, Hugo
Publisher: name: London : Biomed Central
Year: 2009
Subjects: subject: Biochemical properties
subject: Biological activities
subject: Biomedical applications
subject: Chemical precursors
subject: Co-transfections
subject: DNA expressions
subject: DNA molecules
subject: Electron-accepting
subject: Fusion proteins
subject: Gold nanoparticle
subject: Gold Nanoparticles
subject: Mammalian cells
subject: Plasma membranes
subject: Plasmid DNA
subject: Positively charged
subject: Protein accumulation
subject: Transfection efficiency
subject: Transfection reagents
subject: Ultrashort-pulsed laser
subject: Bioactivity
subject: Cell membranes
subject: DNA
subject: Genes
subject: Laser ablation
subject: Lasers
subject: Mammals
subject: Molecular biology
subject: Nanoparticles
subject: Pulsed laser applications
subject: Transcription factors
subject: Nucleic acids
subject: enhanced green fluorescent protein
subject: gold nanoparticle
subject: high mobility group B1 protein
subject: hybrid protein
subject: plasmid DNA
subject: plasmid vector
subject: recombinant protein
subject: animal cell
subject: article
subject: cell nucleus
subject: controlled study
subject: gene expression
subject: genetic transfection
subject: laser
subject: nanobiotechnology
subject: nonhuman
subject: particle size
subject: protein localization
subject: protein transport
subject: recombinant plasmid
subject: Mammalia
subject: Dewey Decimal Classification::500 | Naturwissenschaften::540 | Chemie
subject: Dewey Decimal Classification::600 | Technik::610 | Medizin, Gesundheit

description: Ultrashort pulsed laser ablation in liquids represents a powerful tool for the generation of pure gold nanoparticles (AuNPs) avoiding chemical precursors and thereby making them especially interesting for biomedical applications. However, because of their electron accepting properties, laser-generated AuNPs might affect biochemical properties of biomolecules, which often adsorb onto the nanoparticles. We investigated possible effects of such laser-generated AuNPs on biological functionality of DNA molecules. We tested four differently sized and positively charged AuNPs by incubating them with recombinant eGFP-C1-HMGB1 DNA expression plasmids that code for eGFP fusion proteins and contain the canine architectural transcription factor HMGB1. We were able to show that successfully transfected mammalian cells are still able to synthesize and process the fusion proteins. Our observations revealed that incubation of AuNP with the plasmid DNA encoding the recombinant canine HMGB1 neither prevented the mediated uptake of the vector through the plasma membrane in presence of a transfection reagent nor had any effect on the transport of the synthesized fusion proteins to the nuclei. Biological activity of the recombinant GFP-HMGB1 fusion protein appears to have not been affected either, as a strong characteristic protein accumulation in the nucleus could be observed. We also discovered that transfection efficiencies depend on the size of AuNP. In conclusion, our data indicate that laser-generated AuNPs present a good alternative to chemically synthesized nanoparticles for use in biomedical applications.