scholarly journals Efficient and Low Cytotoxicity Gene Carriers Based on Amine-Functionalized Polyvinylpyrrolidone

Polymers ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 2724
Author(s):  
Anselmo Del Prado ◽  
Ana Civantos ◽  
Enrique Martínez-Campos ◽  
Pavel A. Levkin ◽  
Helmut Reinecke ◽  
...  
Keyword(s):  
Viral Vectors ◽  
Transfection Efficiency ◽  
Positive Control ◽  
Viral Gene ◽  
General Tendency ◽  
Molar Ratios ◽  
Gene Carriers ◽  
Low Cytotoxicity ◽  
Average Size

Non-viral vectors are a safety tool for gene therapy to deliver therapeutic genes. Among the different non-viral vectors, polyvinylpyrrolidone (PVP), a well-known hydrosoluble, neutral, and non-toxic polymer, satisfies the requirements and becomes a suitable candidate for gene delivery. In this study, we describe the preparation of polyvinylpyrrolidones decorated with pyrrolidine, piperidine, and piperazine groups, and evaluate them in vitro as non-viral gene carriers. The properties of these new systems are compared with those of hyperbranched polyethyleneimine (PEI) used as a positive control. Their ability to complex DNA at different N/P molar ratios, from 1:1 up to 10:1, was studied through agarose gel electrophoresis and dynamic light scattering. The resulting complexes (polyplexes) were characterized and evaluated in vitro with murine fibroblast (Swiss 3T3) as non-viral gene carriers, using luciferase as the reporter gene and a calcein cytocompatibility assay. All the copolymers condensed DNA to a particle average size between 100–400 nm when used at N/P ratios of 4:1 or higher. The copolymers with piperidine groups showed higher transfection efficiency than the pyrrolidine and piperazine modified copolymers, and even higher than the positive control of PEI at N/P ratios of 4:1 or higher. All the synthesized polyplexes from an aminated PVP displayed a general tendency of high cytocompatibility (75–95%) in comparison with the positive control PEI (55%).

scholarly journals TAMI-14. NANOPARTICLE DELIVERY OF PD-L1 CRISPR/CAS9 PLASMID DNA FOR ANTI-GLIOBLASTOMA IMMUNOTHERAPY

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii216-ii216
Author(s):  
Javier Fierro ◽  
An Tran ◽  
Chris Factoriza ◽  
Brandon Chin ◽  
Huanyu Dou
Keyword(s):  
Plasmid Dna ◽  
Viral Vectors ◽  
New Technologies ◽  
Transfection Efficiency ◽  
Guide Rna ◽  
Immune Balance ◽  
Main Challenge ◽  
Low Cytotoxicity

Abstract Glioblastoma multiforme (GBM) is a devastating cancer that develops from astrocytes in the brain. GBM is fast acting and kills 90% of patients within 5 years. Several immunotherapies have been developed to treat GBM, however, major challenges still persist. For example, checkpoint proteins such as programmed cell death protein 1 (PD-1) and its ligand, programmed death ligand 1 (PD-L1), are upregulated in GBM cells to evade the immune system. Targeting PD-L1 for genetic knockdown is thus a promising avenue for the treatment of GBM. However, PD-L1 protein inhibitors have been shown to cause immune overreaction and toxicity, therefore requiring new technologies. CRISPR/Cas9 gene editing has been widely used for the study and treatment of many diseases, but has not been extensively studied for the treatment of GBM. The main challenge is developing a gene delivery platform for the delivery of CRISPR/Cas9 plasmid DNA (pDNA). Many viral vectors have been used for the delivery of pDNA, but unfortunately are associated with high toxicity. Nanotechnology is emerging as a new platform for the delivery of pDNA as it shows high transfection efficiency with low cytotoxicity. We developed a cationic core-shell nanoparticle (NP) capable of carrying CRISPR/Cas9 pDNA. This plasmid contains multiple guide RNA (gRNA) expression cassettes for the knockdown of PD-L1. PDL1gRNA-CRISPR/Cas9pDNA-NPs were taken up by U87 cells within 30 minutes, and entered into the nucleus at 2 hours. The effective delivery of PDL1gRNA-CRISPR/Cas9pDNA-NPs led to the expression of PD-L1 gRNA and Cas9 enzyme, and the knockdown of PD-L1. Regulation of immune balance was determined after PD-L1 knockdown in vitro and in vivo. Our study shows the potential of NP-based PDL1gRNA-CRISPR/Cas9 delivery as an anti-GBM immunotherapy for clinical applications.

DEGRADABLE POLYETHYLENIMINE DERIVATIVES AS GENE CARRIERS

Nano LIFE ◽  
2012 ◽  
Vol 02 (01) ◽  
pp. 1230004 ◽  
Author(s):  
YOU-KYOUNG KIM ◽  
QUYNH-PHUONG LUU ◽  
MOHAMMAD ARIFUL ISLAM ◽  
YUN-JAIE CHOI ◽  
CHONG-SU CHO ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Viral Vectors ◽  
Large Scale ◽  
Transfection Efficiency ◽  
Endosomal Escape ◽  
Scale Production ◽  
Nonviral Vectors ◽  
High Transfection Efficiency ◽  
Gene Carriers

Gene therapy is a treatment for inborn and acquired diseases, although the development of safe and effective gene delivery system is a great challenge to make a gene therapy a success. Viral vectors have been used in a majority of clinics because of their high transfection efficiency in vitro and in vivo. However, their use has been limited because of several drawbacks, such as induction of immune response, recombination of wild-type viruses, limitation in the size of inserted gene, and difficulty in large-scale production. Nonviral vectors have been widely proposed safe alternatives to viral vectors because they have low immunogenicity, flexibility in the size of gene to be delivered, cell targetibility, and easy scalability of production, although they have low transfection efficiency compared to viral vectors. Among nonviral vectors, polyethylenimine (PEI) has been widely used as a standard gene carriers due to its high pH-buffering capacity for endosomal escape although high-molecular-weight PEI is too toxic owing to non-degradability. Recently, many types of degradable PEI have been studied due to high transfection efficiency with lower cytotoxicity. This review explains recent progress on the development of degradable PEIs as nonviral vectors. The present paper summarizes the transfection efficiency of DNA or silencing efficiency of small interfering RNA (siRNA) based on the kinds of degradable linkage between low PEI and crosslinkers. Degradable linkages, such as ester, disulfide, imines, carbamate, amide and ketal in the degradable PEIs are covered.

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.

scholarly journals Biodegradable Gene Carriers Containing Rigid Aromatic Linkage with Enhanced DNA Binding and Cell Uptake

Polymers ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 1080 ◽  
Author(s):  
Ju-Hui Zhang ◽  
Hui-Zhen Yang ◽  
Ji Zhang ◽  
Yan-Hong Liu ◽  
Xi He ◽  
...  
Keyword(s):  
Dna Binding ◽  
Laser Scanning ◽  
Transfection Efficiency ◽  
Laser Scanning Microscopy ◽  
Cell Uptake ◽  
Confocal Laser ◽  
Viral Gene ◽  
Cationic Polymers ◽  
Aromatic Rings ◽  
Gene Carriers

The linking and modification of low molecular weight cationic polymers (oligomers) has become an attracted strategy to construct non-viral gene carriers with good transfection efficiency and much reduced cytotoxicity. In this study, PEI 600 Da was linked by biodegradable bridges containing rigid aromatic rings. The introduction of aromatic rings enhanced the DNA-binding ability of the target polymers and also improved the stability of the formed polymer/DNA complexes. The biodegradable property and resulted DNA release were verified by enzyme stimulated gel electrophoresis experiment. These materials have lower molecular weights compared to PEI 25 kDa, but exhibited higher transfection efficiency, especially in the presence of serum. Flow cytometry and confocal laser scanning microscopy results indicate that the polymers with aromatic rings could induce higher cellular uptake. This strategy for the construction of non-viral gene vectors may be applied as an efficient and promising method for gene delivery.

Bio-inspired gene carriers with low cytotoxicity constructed via the assembly of dextran nanogels and nano-coacervates

Nanomedicine ◽  
2020 ◽  
Vol 15 (13) ◽  
pp. 1285-1296 ◽  
Author(s):  
Chenglong Wang ◽  
Jiayi You ◽  
Miaomiao Gao ◽  
Peipei Zhang ◽  
Guoxiong Xu ◽  
...  
Keyword(s):  
Positive Charge ◽  
Transfection Efficiency ◽  
Gene Carrier ◽  
Gene Carriers ◽  
Environmentally Responsive ◽  
Potential Applications ◽  
Internal Peptide ◽  
The Common ◽  
Low Cytotoxicity ◽  
Peptide Gene

Aim: To achieve safe and biocompatible gene carriers. Materials & methods: A core/shell-structured hierarchical carrier with an internal peptide/gene coacervate ‘core’ and a dextran nanogel ‘shell’ on the surface has been designed. Results: The dextran nanogels shield coacervate (DNSC) can effectively condense genes and release them in reducing environments. The dextran nanogel-based ‘shell’ can effectively shield the positive charge of the peptide/gene coacervate ‘core’, thus reducing the side effects of cationic gene carriers. In contrast with the common nonviral gene carriers that had high cytotoxicities, the DNSC showed a high transfection efficiency while maintaining a low cytotoxicity. Conclusion: The DNSC provides an effective environmentally responsive gene carrier with potential applications in the fields of gene therapy and gene carrier development.

Receptor-Mediated Gene Delivery Using Chitosan Derivatives In Vitro and In Vivo

2007 ◽  
Vol 342-343 ◽  
pp. 449-452 ◽  
Author(s):  
Tae Hee Kim ◽  
Hua Jin ◽  
Hyun Woo Kim ◽  
Myung Haing Cho ◽  
Jae Woon Nah ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Delivery System ◽  
Transfection Efficiency ◽  
Viral Gene ◽  
Gene Delivery System ◽  
Targeted Gene Delivery ◽  
Cell Specificity ◽  
Selective Delivery

The key strategy for the advancement of gene therapy is the development of an efficient targeted gene delivery system into cells. The targeted gene delivery system is especially important in non-viral gene transfer which shows the relatively low transfection efficiency. It also opens the possibility of selective delivery of therapeutic plasmids to specific tissues. Chitosan has been considered to be a good candidate for gene delivery system, since it is already known as a biocompatible, biodegradable, and low toxic material with high cationic potential. However, low specificity and low transfection efficiency of chitosan need to be overcome prior to clinical trial. In this study, we focused on the chemical modification of chitosan for enhancement of cell specificity and transfection efficiency. Also, the potential of clinical application was investigated.

scholarly journals The Development of Functional Non-Viral Vectors for Gene Delivery

2019 ◽  
Vol 20 (21) ◽  
pp. 5491 ◽  
Author(s):  
Patil ◽  
Gao ◽  
Lin ◽  
Li ◽  
Dang ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Targeted Delivery ◽  
Viral Vectors ◽  
High Efficiency ◽  
Transfection Efficiency ◽  
Cationic Lipids ◽  
Gene Vectors ◽  
Potential Applications ◽  
Low Toxicity ◽  
Low Cytotoxicity

Gene therapy is manipulation in/of gene expression in specific cells/tissue to treat diseases. This manipulation is carried out by introducing exogenous nucleic acids, such as DNA or RNA, into the cell. Because of their negative charge and considerable larger size, the delivery of these molecules, in general, should be mediated by gene vectors. Non-viral vectors, as promising delivery systems, have received considerable attention due to their low cytotoxicity and non-immunogenicity. As research continued, more and more functional non-viral vectors have emerged. They not only have the ability to deliver a gene into the cells but also have other functions, such as the performance of fluorescence imaging, which aids in monitoring their progress, targeted delivery, and biodegradation. Recently, many reviews related to non-viral vectors, such as polymers and cationic lipids, have been reported. However, there are few reviews regarding functional non-viral vectors. This review summarizes the common functional non-viral vectors developed in the last ten years and their potential applications in the future. The transfection efficiency and the transport mechanism of these materials were also discussed in detail. We hope that this review can help researchers design more new high-efficiency and low-toxicity multifunctional non-viral vectors, and further accelerate the progress of gene therapy.

In vitro and in vivo gene transfection using biodegradable and low cytotoxic nanomicelles based on dendritic block copolymers

2015 ◽  
Vol 3 (4) ◽  
pp. 688-699 ◽  
Author(s):  
Yan Liu ◽  
Chao Lin ◽  
Jianbo Li ◽  
Yang Qu ◽  
Jie Ren
Keyword(s):  
Block Copolymers ◽  
Viral Vectors ◽  
Gene Transfection ◽  
Low Cytotoxicity

Dendritic PCL-b-PDMAEMA copolymers have been used as non-viral vectors for gene transfection and exhibited high transfection efficiencies and low cytotoxicity.

Defined pH-sensitive nanogels as gene delivery platform for siRNA mediated in vitro gene silencing

2017 ◽  
Vol 5 (11) ◽  
pp. 2328-2336 ◽  
Author(s):  
Mathias Dimde ◽  
Falko Neumann ◽  
Felix Reisbeck ◽  
Svenja Ehrmann ◽  
Jose Luis Cuellar-Camacho ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Gene Silencing ◽  
Transfection Efficiency ◽  
Genetic Material ◽  
Ph Sensitive ◽  
Carrier System ◽  
High Transfection Efficiency ◽  
Delivery Platform ◽  
Low Cytotoxicity

An advanced cationic carrier system which combines high transfection efficiency with low cytotoxicity and a control over the release of the encapsulated genetic material by the reduction of the multivalent architecture upon pH triggered degradation was developed.

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