Electrosprayed chitosan nanoparticles: facile and efficient approach for bacterial transformation

Abstract

A rapid and efficient procedure for DNA transformation is a key prerequisite for successful cloning and genomic studies. While there are efforts to develop a facile method, so far obtained efficiencies for alternative methods have been unsatisfactory (i.e. 10 5-10 6 CFU/μg plasmid) compared with conventional method (up to 10 8 CFU/μg plasmid). In this work, for the first time, we prepared chitosan/pDNA nanoparticles by electrospraying methods to improve transformation process. Electrospray method was used for chitosan/pDNA nanoparticles production to investigate the non-competent bacterial transformation efficiency; besides, the effect of chitosan molecular weight, N/P ratio and nanoparticle size on non-competent bacterial transformation efficiency was evaluated too. The results showed that transformation efficiency increased with decreasing the molecular weight, N/P ratio and nanoparticles size. In addition, transformation efficiency of 1.7 × 10 8 CFU/μg plasmid was obtained with chitosan molecular weight, N/P ratio and nanoparticles size values of 30 kDa, 1 and 125 nm. Chitosan/ pDNA electrosprayed nanoparticles were produced and the effect of molecular weight, N/P and size of nanoparticles on transformation efficiency was evaluated. In total, we present a facile and rapid method for bacterial transformation, which has comparable efficiency with the common method.

Electrospraying: Possibilities and Challenges of Engineering Carriers for Biomedical Applications—A Mini Review

“…To transform bacterial cells with exogenous DNA, chemo-transformation and electroporation are commonly used. However, these methods are complex and expensive, and the step of preparing competent cells is time-consuming (Abyadeh et al, 2017). Electrospraying of DNA plasmid (pDNA) in a suitable buffer (such as high-conductivity culture medium) had been suggested as an alternative for bacterial/cell transformation with high efficiency (Boda et al, 2018).…”

Recent Progress of Chitosan and Chitosan Derivatives-Based Nanoparticles: Pharmaceutical Perspectives of Oral Insulin Delivery

“…Additionally, material-related parameters are: type of polymer, its MW and concentration, as well as excipients used [ 82 ]. Chitosan nanoparticles have been recently fabricated using electrospraying technique, exhibited unique features that may have wide applications in pharmaceutical and biomedical fields ( Figure 8 ) [ 83 , 84 , 85 , 86 ]. The advantages of this method can be summarised by its simple and one step technique, high MW polymers can be used, monodispersed nanoparticles can be obtained and low cost [ 87 ], however, the disadvantage is associated with the low yield product.…”

Towards Building a Plant Cell Atlas

“…A controllable promoter to titrate expression, such as a steroid-inducible promoter, could mitigate the overexpression problem. Exciting new methods have been developed using nanoparticles to aid cellular transformation, including electrospray [13], magnetofection [14], and nanostraw-mediated electroporation [15]. If adapted to plant tissue, these techniques may allow large-scale transient transformation of probes into differentiated plant tissue, and even extend studies to a wide range of species with specialized cell types.…”