scholarly journals Highly Effective Crosslinker for Redox-Sensitive Gene Carriers

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Lin Lin ◽  
Jie Chen ◽  
Yingying Hu ◽  
Huapan Fang ◽  
Kui Wang ◽  
...  
Keyword(s):  
Room Temperature ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Binary Complex ◽  
Cell Internalization ◽  
Highly Effective ◽  
High Gene ◽  
High Level ◽  
In Vitro Transfection

Polyethyleneimine (PEI) has been extensively used as a common gene carrier due to its high gene transfection efficiency. PEI1.8k shows significantly lower cytotoxicity than its high molecular weight counterparts. However, it also has the problem of low gene transfection efficiency. To address the dilemma, a highly effective crosslinker (DTME) was synthesized to react with PEI1.8k to obtain CS-PEI1.8k. The reaction showed several advantages, such as a fast process in room temperature within nine hours with the product which can directly complex with DNA after removing the solvent. The ability of CS-PEI1.8k to agglomerate with DNA was proven by particle size, zeta potential, and gel retardation assays. The cytotoxic in vitro transfection ability and cell internalization capacity of CS-PEI1.8k were tested to verify the transfection capacity of CS-PEI1.8k. Moreover, we also studied the mechanism of the relatively high level of gene transfection by this binary complex compared with PEI25k.

scholarly journals Development of Lyophilized Gemini Surfactant-Based Gene Delivery Systems: Influence of Lyophilization on the Structure, Activity and Stability of the Lipoplexes

2012 ◽  
Vol 15 (4) ◽  
pp. 548 ◽  
Author(s):  
Waleed Mohammed-Saeid ◽  
Deborah Michel ◽  
Anas El-Aneed ◽  
Ronald E Verrall ◽  
Nicholas H Low ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Gemini Surfactant ◽  
Gemini Surfactants ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Physiochemical Properties ◽  
Cationic Gemini Surfactants ◽  
The Stability ◽  
In Vitro Transfection

Purpose. Cationic gemini surfactants have been studied as non-viral vectors for gene therapy. Clinical applications of cationic lipid/DNA lipoplexes are restricted by their instability in aqueous formulations. In this work, we investigated the influence of lyophilization on the essential physiochemical properties and in vitro transfection of gemini surfactant-lipoplexes. Additionally, we evaluated the feasibility of lyophilization as a technique for preparing lipoplexes with long term stability. Methods. A gemini surfactant [12-7NH-12] and plasmid DNA encoding for interferon-γ were used to prepare gemini surfactant/pDNA [P/G] lipoplexes. Helper lipid DOPE [L] was incorporated in all formulation producing a [P/G/L] system. Sucrose and trehalose were utilized as stabilizing agents. To evaluate the ability of lyophilization to improve the stability of gemini surfactant-based lipoplexes, four lyophilized formulations were stored at 25˚C for three months. The formulations were analyzed at different time-points for physiochemical properties and in vitro transfection. Results. The results showed that both sucrose and trehalose provided anticipated stabilizing effect. The transfection efficiency of the lipoplexes increased 2-3 fold compared to fresh formulations upon lyophilization. This effect can be attributed to the improvement of DNA compaction and changes in the lipoplex morphology due to the lyophilization/rehydration cycles. The physiochemical properties of the lyophilized formulations were maintained throughout the stability study. All lyophilized formulations showed a significant loss of gene transfection activity after three months of storage. Nevertheless, no significant losses of transfection efficiency were observed for three formulations after two months storage at 25 ˚C. Conclusion. Lyophilization significantly improved the physical stability of gemini surfactant-based lipoplexes compared to liquid formulations. As well, lyophilization improved the transfection efficiency of the lipoplexes. The loss of transfection activity upon storage is most probably due to the conformational changes in the supramolecular structure of the lipoplexes as a function of time and temperature rather than to DNA degradation. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

Continuous Vector-free Gene Transfer with a Novel Microfluidic Chip and Nanoneedle Array

2018 ◽  
Vol 16 (2) ◽  
pp. 164-170 ◽  
Author(s):  
Dong Huang ◽  
Deyao Zhao ◽  
Jinhui Li ◽  
Yuting Wu ◽  
Lili Du ◽  
...  
Keyword(s):  
Microfluidic Chip ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Basic Research ◽  
Nanoneedle Array ◽  
Contact Frequency ◽  
Delivery Platform ◽  
High Gene ◽  
Free Gene

Background: Delivery of foreign cargoes into cells is of great value for bioengineering research and therapeutic applications. Objective: In this study, we proposed and established a carrier-free gene delivery platform utilizing staggered herringbone channel and silicon nanoneedle array, to achieve high-throughput in vitro gene transfection. </P><P> Methods: With this microchip, fluidic micro vortices could be induced by the staggered-herringboneshaped grooves within the channel, which increased the contact frequency of the cells with the channel substrate. Transient disruptions on the cell membrane were well established by the nanoneedle array on the substrate. </P><P> Result: Compared to the conventional nanoneedle-based delivery system, proposed microfluidic chip achieved flow-through treatment with high gene transfection efficiency (higher than 20%) and ideal cell viability (higher than 95%). It provides a continuous processing environment that can satisfy the transfection requirement of large amounts of biological molecules, showing high potential and promising prospect for both basic research and clinical application.

Synthesis and Characterization of Polyethylenimine-Graft-Poly(L-Lactide-Co-Glycolide) Block Copolymers for Gene Delivery

2007 ◽  
Vol 342-343 ◽  
pp. 521-524
Author(s):  
Oju Jeon ◽  
Su Jin Song ◽  
Min Hyung Lee ◽  
Sang Woo Seo ◽  
Cha Yong Choi ◽  
...  
Keyword(s):  
Block Copolymers ◽  
Gene Delivery ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Gel Permeation Chromatography ◽  
Copolymer Composition ◽  
High Gene ◽  
Good Biocompatibility ◽  
Delivery Vehicles

Polyethylenimine-graft-poly(L-lactide-co-glycolide) (PEI-g-PLGA) block copolymers were prepared by a ring-opening polymerization of L-lactide and glycolide using PEI as a macroinitiator and stannous octoate as a catalyst in dimethylformamide at 100 °C. The molecular structure of the block copolymers was evaluated with 1H-NMR, and the molecular weight of the block copolymers was determined with gel permeation chromatography. The thermal properties were investigated using differential scanning calorimetery and thermogravimetric analysis. The zetapotential of the pDNA/copolymer complexes was evaluated with dynamic laser light scattering. Cytotoxicity and gene transfection efficiency of PEI-g-PLGA were tested in vitro using human embryonic kidney 293 cell culture. The pDNA/copolymer complexes (N/P = 10) showed a lower zeta-potential than pDNA/PEI25kDa complex, suggesting the lower toxicity of the pDNA/copolymer complexes. The copolymer composition was found to significantly affect the gene transfection efficiency of the pDNA/copolymer complexes. The copolymers with lower contents of PLGA showed higher gene transfection efficiency. These results indicate that these block copolymers are promising candidates for gene delivery vehicles, featuring good biocompatibility, potential biodegradability, and relatively high gene transfection efficiency.

Non-Viral Vectors for Gene Delivery

2018 ◽  
Vol 9 (1) ◽  
pp. 4-11 ◽  
Author(s):  
Aparna Bansal ◽  
Himanshu
Keyword(s):  
Gene Therapy ◽  
Viral Vectors ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Expression System ◽  
Target Cells ◽  
Specific Expression ◽  
Naked Dna

Introduction: Gene therapy has emerged out as a promising therapeutic pave for the treatment of genetic and acquired diseases. Gene transfection into target cells using naked DNA is a simple and safe approach which has been further improved by combining vectors or gene carriers. Both viral and non-viral approaches have achieved a milestone to establish this technique, but non-viral approaches have attained a significant attention because of their favourable properties like less immunotoxicity and biosafety, easy to produce with versatile surface modifications, etc. Literature is rich in evidences which revealed that undoubtedly, non–viral vectors have acquired a unique place in gene therapy but still there are number of challenges which are to be overcome to increase their effectiveness and prove them ideal gene vectors. Conclusion: To date, tissue specific expression, long lasting gene expression system, enhanced gene transfection efficiency has been achieved with improvement in delivery methods using non-viral vectors. This review mainly summarizes the various physical and chemical methods for gene transfer in vitro and in vivo.

Retracted Article: A multifunctional self-dissociative polyethyleneimine derivative coating polymer for enhancing the gene transfection efficiency of DNA/polyethyleneimine polyplexes in vitro and in vivo

2015 ◽  
Vol 6 (5) ◽  
pp. 780-796 ◽  
Author(s):  
Cheng Wang ◽  
Xiuli Bao ◽  
Xuefang Ding ◽  
Yang Ding ◽  
Sarra Abbad ◽  
...  
Keyword(s):  
Transfection Efficiency ◽  
Gene Transfection ◽  
Retracted Article ◽  
First Time

A novel coating polymer LPHF is developed for the first time to elevate the transfection efficiency of DP binary polyplexes in vitro and in vivo.

scholarly journals Exploitation of De Novo Helper-Lipids for Effective Gene Delivery.

2009 ◽  
Vol 11 (4) ◽  
pp. 56 ◽  
Author(s):  
Tomoaki Kurosaki ◽  
Takashi Kitahara ◽  
Mugen Teshima ◽  
Koyo Nishida ◽  
Junzo Nakamura ◽  
...  
Keyword(s):  
Gene Delivery ◽  
De Novo ◽  
Transfection Efficiency ◽  
The Other ◽  
Penetration Enhancers ◽  
Cell Toxicity ◽  
In Vivo Transfection ◽  
In Vitro Transfection

Purpose: In gene delivery, a fusogenic lipid such as dioleyl phosphatidylethanolamine (DOPE) which is a component of cationic liposomal vector is important factor for effective transfection efficiency. We investigated the effect of penetration enhancers as alternative helper-lipids to DOPE. Methods: Transdermal penetraion enhancers such as N-lauroylsarcosine (LS), (R)-(+)-limonene (LM), vitamin E (VE), and phosphatidyl choline from eggs (EggPC) were used in this experiments as helper-lipids with N-[1-(2, 3-dioleyloxy) propyl]-N, N, N-trimethlylammonium chloride (DOTMA) and cholesterol (CHOL). We examined in vitro transfection efficiency, cytotoxicity, hematotoxicity, and in vivo transfection efficiency of plasmid DNA/cationic liposomes complexes. Results: In transfection experiments in vitro, the cationic lipoplexes containing LS had highest transfection efficiency among the other lipoplexes independently of FBS. Furthermore, the lipoplexes containing LS had lowest cell toxicity among the other lipoplexes in the presence of FBS. As the results of erythrocytes interaction experiment, DOTMA/LS/CHOL, DOTMA/VE/CHOL, and DOTMA/EggPC/CHOL lipoplexes showed extremely lower hematotoxicity. On the basis of these results, the in vivo transfection efficiencies of the lipoplexes were examined. The lipoplexes containing LS had the highest transfection activity among the other lipoplexes. Conclusion: In conclusion, several transdermal penetration enhancers are available for alternative helper-lipids to DOPE in cationic liposomal vectors. Among them, DOTMA/LS/CHOL lipoplexes showed superior characteristics in in vitro transfection efficiency, cell toxicity, hematotoxicity, and in vivo transfection efficiency.

A general strategy to achieve ultra-high gene transfection efficiency using lipid-nanoparticle composites

Biomaterials ◽  
2014 ◽  
Vol 35 (28) ◽  
pp. 8261-8272 ◽  
Author(s):  
Raviraj Vankayala ◽  
Chi-Shiun Chiang ◽  
Jui-I. Chao ◽  
Chiun-Jye Yuan ◽  
Shyr-Yeu Lin ◽  
...  
Keyword(s):  
Transfection Efficiency ◽  
Gene Transfection ◽  
General Strategy ◽  
Lipid Nanoparticle ◽  
High Gene ◽  
Nanoparticle Composites

Autocrine IL6/STAT3 Signaling Enhances Survival of Chronic Lymphocytic Leukaemia Cells

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3598-3598
Author(s):  
Fengting Liu ◽  
Li Jia ◽  
Timothy Farren ◽  
John G. Gribben ◽  
Samir Agrawal
Keyword(s):  
Gene Transfection ◽  
Fluorescent Microscopy ◽  
Ros Generation ◽  
Apoptosis Resistance ◽  
Cytochrome C Release ◽  
Cancer Cell Survival ◽  
High Level ◽  
Stat3 Signalling ◽  
Microscopy Techniques

Abstract Abstract 3598 Signal transducer and activator of transcription 3 (STAT3) proteins have been found to play an important role in cancer cell survival and proliferation. The activation of STAT3 signalling pathways require interleukin-6 (IL6) and tyrosine kinase (JAK2) phosphorylation. Previous studies have shown that in patients with CLL elevated levels of serum IL6 correlate with adverse disease features and shorter survival. Methods: We investigated the relationship of CLL cell autocrine IL6 production, STAT3 activation and apoptosis resistance. Thirty-six CLL patients with high lymphocyte counts were investigated. Westen blot, flow cytometry, gene transfection and fluorescent microscopy techniques were used. Results: Autocrine IL6 production in CLL cells cultured for 24 hours was higher in Binet stage B/C patients (61pg/ml, range 1.4–297pg/ml) as compared with stage A patients (6.1pg/ml, range 0–23pg/ml) (p=0.02). Patients with high autocrine IL6 production had a higher ratio of phosphorylated STAT3/ total STAT3, indicating a high level of STAT3 activation and apoptosis resistance. Activation of the IL6/JAK2/STAT3 pathway by exogenous IL6 led to increased expression of anti-apoptotic proteins Mcl-1 and Bcl-xl. STAT3 activation by exogenous IL6 led to increased resistance of CLL cells to spontaneous in vitro apoptosis (mean increase in viable cells over control: 15%, range 2–36%; p=0.00008). This was associated with preservation of mitochondrial function: decreased cytochrome C release (p=0.004), mitochondrial membrane potential collapse (p=0.0004) and ROS generation (p=0.0007). This study demonstrates that higher autocrine IL6 production by CLL correlates with STAT3 activation and apoptosis resistance in CLL. The IL6/JAK2/STAT3 signal pathway may reveal new therapeutic targets. Disclosures: Gribben: Roche: Consultancy; Celgene: Consultancy; GSK: Honoraria; Napp: Honoraria.

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