title: Nanoparticles facilitate gene delivery to microorganisms via an electrospray process.
abstract: In this study, we developed a technique for delivering genes to microorganisms via electrospray of gold nanoparticles. During the electrospray process, charged monodisperse nano-droplets (a mixture of pET30a-GFP plasmid and nano-sized gold particles) were accelerated and deposited on a thin layer of non-competent Escherichia coli cells. Via antibiotic selection, transformed cells containing green fluorescent protein appeared on the agar plates. PCR amplification and restriction enzyme analysis further confirmed that pET30a-GFP plasmid had successfully been delivered into the non-competent E. coli cells. The transformation efficiencies were optimized under different electrospray conditions. Among several electrospray buffer solutions, CaCl(2) (0.01M) was found to be the best for gene delivery. Furthermore, gold nanoparticles (NPs, 50 nm diameter) significantly improved plasmid transformation efficiency by 5-7 fold (up to 2×10(6) CFU/μg plasmid) compared with that obtained using naked plasmid. Electronic microscopy images and gel electrophoresis showed that the morphology of plasmids remained unchanged during the electrospray process, but cellular membrane integrity was reduced after being electrosprayed with gold NPs and CaCl(2) buffer solutions. This gene delivery method has the potential to work for many other microorganisms.
authors.affiliation: Department of Energy, Environmental, and Chemical Engineering, Washington University, St. Louis, MO 63130, USA.
authors.forename: Yi-Hsuan
authors.initials: YH
authors.lastname: Lee
authors.forename: Bing
authors.initials: B
authors.lastname: Wu
authors.forename: Wei-Qin
authors.initials: WQ
authors.lastname: Zhuang
authors.forename: Da-Ren
authors.initials: DR
authors.lastname: Chen
authors.forename: Yinjie J
authors.initials: YJ
authors.lastname: Tang
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005827:Genetics, Microbial / Q000379:methods*; D006046:Gold; D053758:Nanoparticles; D010957:Plasmids / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial; D006046:Gold