Ternary complex of plasmid DNA with NLS-Mu-Mu protein and cationic niosome for biocompatible and efficient gene delivery: a comparative study with protamine and lipofectamine

Abstract

Non-viral gene delivery methods are considered due to safety and simplicity in human gene therapy. Since the use of cationic peptide and niosome represent a promising approach for gene delivery purposes we used recombinant fusion protein and cationic niosome as a gene carrier. A multi-domain fusion protein including nuclear localization motif (NLS) and two DNA-binding (Mu) domains, namely NLS-Mu-Mu (NMM) has been designed, cloned and expressed in E. coli DE3 strain. Afterward, the interested protein was purified by affinity chromatography. Binary vectors based on protein/DNA and ternary vectors based on protein/DNA/niosome were prepared. Protamine was used as a control. DNA condensing properties of NMM and protamine were evaluated by various experiments. Furthermore, we examined cytotoxicity, hemolysis and transfection potential of the binary and ternary complexes in HEK293T and MCF-7 cell lines. Protamine and Lipofectamine™2000 were used as positive controls, correspondingly. The recombinant NMM was expressed and purified successfully and DNA was condensed efficiently at charge ratios that were not harmful to cells. Peptidoplexes showed transfection efficiency (TE) but ternary complexes had higher TE. Additionally, NMM ternary complex was more efficient compared to protamine ternary vectors. Our results showed that niosomal ternary vector of NMM is a promising non-viral gene carrier to achieve an effective and safe carrier system for gene therapy.

Poly-l-lysine-coated superparamagnetic nanoparticles: a novel method for the transfection of pro-BDNF into neural stem cells

“…A, The percentage of viable cells (NSCs) loaded with supra paramagnetic iron oxide NPs (SPIONs). The concentrations of SPIONs as indicated (10,25,50, and 100 mg/mL) at different incubation times (1, 1.5, 3, 6, 12, and 24 hr, respectively). B, the percentage of viable cells loaded with SPIONs coated with ploy-L-Lysine (SPIONs-PLL) at 25, 50, and 100 mg/mL.…”

“…Silica nanoparticles (NPs) have been demonstrated for achieving the impermanent gene transfer in vivo as a non-viral vector systems [9]. Some of non-viral carriers were designed and developed to overcome the challenges of ineffective DNA-endosomal capture and release from the carrier vector into the nucleus, and represent a promising methods for gene delivery purposes [10]. Superparamagnetic iron oxide NPs (SPIONs) are widely used for biomedical applications, such as target gene delivery systems and gene therapy, It can be used also in cancer chemotherapy as drug carrier after coated with biocompatible polymers [11,12] SPIONs with unique magnetic properties are being actively investigated as magnetic resonance imaging contrast agents [12], It can be properly designed and engineered to reach their target tissue with less nonspecific cellular interactions [13,14].…”

Development and characterization of a novel cationic PEGylated niosome-encapsulated forms of doxorubicin, quercetin and siRNA for the treatment of cancer by using combination therapy

“…Niosome, a bilayer non-ionic surfactant-based vesicle, is a novel drug delivery system. Having the nanometric scale size, biodegradability, relatively nontoxic property, remarkable stability, long storage time and low cost has enabled niosomes to be forerunners of other nano-carriers [15]. According to Figure 1, when the present delivery system is accompanied with the external stimuli such as pH and the temperature, it triggers the release of the drugs named thermo-and pH-sensitive niosome.…”

A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation

“…Interestingly, after electrotransfection of GFP-vinculin plasmid, we further delivered phalloidin (a fluorescent F-actin probe) into the fibroblasts, demonstrating the capability of our setup in delivering multiple dyes/particles into live cells in a controllable manner, which can greatly help researchers in elucidating the individual and collective roles of different proteins in various cellular processes. We also want to point out that, compared to conventional methods such as bulk electroporation [29] (requiring high-operation voltage, typically of the order of hundreds of volts) and rather expensive transfection reagents like lipofectamine [30], our micro-electroporation approach could offer advantages such as easy/safe to operate, low cost and real-time monitoring.…”