Introduction to Non Viral Vectors

Non-viral vectors, such as lipid nanoparticles, have attracted significant interest, mainly due to their successful use in vaccines in the current COVID-19 pandemic.

In the present study, hBMSCs were used as gene therapy non-viral vectors of miR-451 to treat glioma cells in vitro and in vivo.

Non-viral vectors may represent an alternative, however, reports analyzing their immunogenicity are still lacking.

We discuss gene delivery systems including viral and non-viral vectors as well as gene editing techniques, mainly CRISPR-Cas9, and epigenetic treatments including antisense and siRNA therapeutics.

Integrative non-viral vectors such as transposons engineered to mediate targeted gene transfer into safe harbor sites in the genome may be a promising approach for the production of therapeutic proteins or for gene therapy in an efficient and secure way.

To avoid these drawbacks and ensure efficient delivery to target cells, viral and non-viral vectors are the two main approaches currently employed.

Here, we have developed novel ferrocene-conjugated cationic tocopheryl aggregates as non-viral vectors.

Various viral and non-viral vectors have been utilized for gene editing to intervene gene expression patterns during the cardiac remodeling process.

Non-viral vectors offer the potential to deliver nucleic acids including mRNA and DNA into cells in vivo.

Even then, such treatment has mainly entailed ex-vivo strategies, where gene edited cells are introduced into the patients’ bodies rather than in-vivo strategies, where the gene-editing tools are administered themselves instead of the altered cells using viral or non-viral vectors [4].

Consequently, the development of non-viral vectors has gained attention, and nowadays, gene delivery mediated by these systems is considered as the cornerstone of modern gene therapy due to relevant advantages such as low toxicity, poor immunogenicity and high packing capacity.

Non-viral vectors have been a fast-paced research topic in gene delivery, such as polymers, lipids, inorganic particles, and combinations of different types.

In view of this, the present review article summarizes nanoscale-based adjuvants and delivery vehicles such as viral vectors, virus-like particles and virosomes; non-viral vectors namely nanoemulsions, lipid nanocarriers, biodegradable and non-degradable nanoparticles, calcium phosphate nanoparticles, colloidally stable nanoparticles, proteosomes; and pattern recognition receptors covering c-type lectin receptors and toll-like receptors.

The gene therapy is mediated by the use of viral and non-viral vectors to transport foreign genes into somatic cells to restorative defective genes.

Over the last decade, non-viral vectors have attracted increasing attention owing to their ability to deliver and/or codeliver different cargos for gene therapy (DNA, siRNA, mRNA, etc), as well as their ease of manufacturing, mild immunogenicity, relatively low toxicity, and compatibility with repeated dosing.

Non-viral vectors and their transfection reagents are useful as safe tools for transfection.

Finally, we highlight future perspectives of the CRISPR-Cas9 gene-editing tool and the prospects of using non-viral vectors to improve its bioavailability and therapeutic potential.

In this context, beside other non-viral vectors, cell-penetrating peptides (CPPs) gain more and more interest as delivery systems by forming a variety of nanocomplexes depending on the formulation conditions and the properties of the used CPPs/NAs.

Herein, we review the recent progress towards CRISPR/Cas9 system delivery based on viral and non-viral vectors.

Non-viral vectors are much less successful than viral vectors, but because of their low immune responses and their broad therapeutic DNA ability, they have advantages.

Lipid nanoparticles (LNPs) have been extensively studied as non-viral vectors for the delivery of mRNA to target cells because of their relatively easy and scalable manufacturing processes.

This review discusses these challenges, covering the strategies used to ensure drug stability and to develop formulations involving viral or non-viral vectors, as well as accelerated regulatory approval pathways for gene therapy products.

Nowadays, gene therapy is employed through both viral and non-viral vectors.

This article mainly focuses on the delivering modes of genetic materials in the CNS, which includes viral and non-viral vectors and their application in gene therapy.

Two non-viral vectors, chitosan (Ch) and polyethyleneimine (PEI), were tested to deliver plasmids encoding for vascular endothelial growth factor (pVEGF) and fibroblast growth factor-2 (pFGF2) to human dermal fibroblasts (hDFbs).

Background/Aim: Genetic manipulation of stem cells using non-viral vectors is still limited due to low transfection efficiency.

Several nanotechnologies based non-viral vectors such as liposome, micelles, nanoparticles, dendrimers, exosomes, nanorods and nanobubbles etc.

The top-cited articles on gene delivery were focused on several novel non-viral vectors due to their specialty compared with viral vectors.

Within non-viral vectors, polymeric nanocarriers have offered an ideal platform to be applied as gene delivery systems.

Metal nanoparticles are the most studied objects for medicine, including their application as non-viral vectors.

Non-viral vectors for gene-based therapy.

In comparison with viral vectors, non-viral vectors became an attractive tool due to their reduced potential of biosafety risks, lower cost, higher availability and easy storage.

Although viral vectors have long been the preferred approach to target hepatocytes, an increasing number of non-viral vectors are emerging as highly efficient vehicles for the delivery of genetic material.

Non-viral vectors are widely developed and applied in clinical practice due to their low immunogenicity, good biocompatibility, easy synthesis and modification, and low cost of production.

Nanomaterials are considered as non-viral vectors for gene therapy use.

Over the past few years, attempts have been made to generate highly functional non-viral vectors that display long-term maintenance within cells and promote more sustained gene expression relative to conventional plasmids.

Antigen-specific lung-resident memory T cells (TRMs) constitute the first line of defense that mediates rapid protection against respiratory pathogens and inspires novel vaccine designs against infectious pandemic threats, yet effective means of inducing TRMs, particularly via non-viral vectors, remain challenging.

Minicircles are non-viral vectors with promising characteristics for application in a variety of fields.

Finally, compact non-viral vectors that can overcome the immunological, physical and anatomical and barriers have been developed.

Although the development of gene delivery systems based on non-viral vectors is advancing, it remains a challenge to deliver plasmid DNA into human blood cells.

Among those non-viral vectors, dendrimers have gained increasing interest due to their unique molecular structure and multivalency.

Along with the use of plasmids and viral vectors as cargo material for genome editing, non-viral vectors have also been considered for delivery purposes.

The low transfection rate of non-viral vectors under physiological conditions is a significant issue.

OBJECTIVES Gene delivery by using non-viral vectors is a multi-step process that poses many challenges, either regarding the extracellular or intracellular media.

The use of gene and drug delivery systems such as viral or non-viral vectors and nano-compounds to enhance CFTR expression and the activity of antimicrobials against chronic pulmonary infections respectively, will also be discussed.

Polyethylene glycol-modified (PEGylated) cationic nanoparticles have been used as non-viral vectors for the in vivo delivery of these nucleic acids.

Non-viral vectors, such as lipid- and polymer-based nanoparticles, cationic polymers, and exosomes, can effectively transfer genes into primary neurons.

Consequently, non-viral vectors are good candidates to increase the lifetime and improve their therapeutic efficacy.

Various viral and non-viral vectors are used as a delivery vehicle for introducing specific miRNA inside the cell.

Trends in changing the paradigm of approaches to therapeutic angiogenesis are noted: from viral vectors to non-viral vectors, from gene transfer to the whole organism to targeted transfer to cells and tissues, from single gene use to combination of genes; from DNA therapy to RNA therapy, from in vivo therapy to ex vivo therapy.

The low transfection rate of non-viral vectors under physiological conditions is a significant issue.

Non-viral vectors are attracting much attention due to their safety and facile production compared to viral vectors.

Gene therapy is evolving rapidly with viral and non-viral vectors that were found to be safe and efficient in preclinical trials.

In particular, among non-viral vectors, gene vectors based on chitosan derivatives are considered a promising gene therapy tool and have been extensively studied due to the biocompatibility, biodegradability, and modifiability of chitosan.

In order to deliver the desired gene at a specific site of action, this therapeutic approach uses vectors divided in two major categories, viral vectors and non-viral vectors, with the latter being represented by chemical delivery agents.

These findings provide guidance for the rational design of silica-based non-viral vectors for efficient gene delivery.

Non-viral vectors ‘polymeric and non-polymeric’ show significant tumour shrinkage in pre-clinical trials and now at early-stage clinical trials.

The development of safe non-viral vectors for the highly efficient delivery of nucleic acids is essential for effective gene therapy, and has been attracting much attention.

These findings provide useful guidance for the rational design of silica-based non-viral vectors for efficient gene delivery applications.

Among the large diversity of non-viral vectors, the cationic polymer polyethylenimine (PEI) plays a prominent role in nucleic acid delivery.

Viral and non-viral vectors are widely used to deliver genetic materials, but each has its respective advantages and shortcomings.

Various types of viral and non-viral vectors are designed to deliver the CRISPR/Cas9 machinery into the desired cell.

Viral vectors for vaccine delivery are challenged by recently reported safety issues like immunogenicity and risk for cancer development, and thus there is a growing need for the development of non-viral vectors.

This review mainly illuminates the current widely used gene therapy drugs, summarizes the viral vectors and non-viral vectors that deliver gene therapy drugs, and sums up the application of gene therapy to treat genetic diseases.

Viral and non-viral vectors have been shown to deliver DNA successfully in vivo, but the sustained expression of CFTR was not adequate.

In this review, we summarize the state-of-the-art non-viral vectors that exploit external stimuli (i.

Purpose: We aim herein to review the currently available methods for gene therapy, which include the utilization of viral/non-viral vectors and how they might serve as therapeutic potentials in regenerative medicine for periodontal and peri-implant tissues.

In this review, we discuss the application of dendrimers as both therapeutic agents and non-viral vectors of chemical agents and genes for HIV treatment.

Safe and efficient non-viral vectors for ocular gene therapy are still unmet clinical desires.

Therefore, the targeted delivery of therapeutics by RGD peptide-based non-viral vectors to tumour vasculature and tumour cells is recognised as a promising approach for treating cancer.

Due to the insecurity of viral vectors, non-viral vectors have gained great attention recently.

In gene delivery, non-viral vectors have become the preferred carrier system for DNA delivery.

Cationic solid-lipid nanoparticles (cSLNs) are considered one of the most promising non-viral vectors for nucleic acid delivery.

Employment of several viral and non-viral vectors has enabled efficient delivery of the CRISPR system to target cells or tissues.

Overall, chitosan–heparin polyplexes showed a significant improvement of gene release inside the cells, transfection, and gene silencing efficiency in vitro, suggesting that this fundamental strategy can further improve the transfection efficiency with application of non-viral vectors.

Clinical relevance of non-viral vectors will therefore depend on the ability to engineer optimized systems for cellular delivery in physiological environments.

Non-viral vectors are relatively non-immunogenic, scalable platforms that have had limited success with DNA delivery to the eye.

Two commonly used non-viral vectors (Lipofectamine 2000 and jet polyethylenimine) were used for the delivery of gene fluorescent protein plasmid in HeLa cancer cells.

This information will help to design more effective cationic surfactants as non-viral vectors for gene therapy.

Cell-penetrating peptides (CPP) are broadly recognized as efficient non-viral vectors for the internalization of compounds such as peptides, oligonucleotides or proteins.

On the other hand, non-viral vectors have less toxicity and immunogenic response and are easier to prepare.

Various studies based on the vector-based system have shown the superiority of the lipid over other non-viral vectors.

Within the field of gene therapy, there is a considerable need for the development of non-viral vectors that are able to compete with the efficiency obtained by viral vectors, while maintaining a good toxicity profile and not inducing an immune response within the body.

, by viral or non-viral vectors) approaches.

Alternatively, non-viral vectors mediated gene delivery has the advantage of low immunogenicity despite showing low transgene expression.

Among them, sugar alcohol-based nanoparticle with abundant surface chemistry, numerous hydroxyl groups, acceptable biocompatibility and biodegradable property are considered as the recent additions to the growing list of non-viral vectors.

Calcium phosphate nanoparticles have received particular attention as non-viral vectors for gene therapy, as factors such as plasmid DNAs, microRNAs (miRNA) and silencing RNA (siRNAs) can be easily incorporated on their surface.

The non-viral vectors based on SLN were able to downregulate the MMP-9 expression in HCE-2 cells via gene silencing, and, consequently, to inhibit cell migration and tube formation.

Non-viral vectors, as promising delivery systems, have received considerable attention due to their low cytotoxicity and non-immunogenicity.

ConclusionThe goal of the research for gene delivery system is to develop the clinically relevant vectors such as viral and non-viral vectors that use to combat elusive diseases such as AIDS, cancer, Alzheimer, etc.

To form bio-inspired non-viral vectors for DNA delivery, the polysaccharide dextran is allowed to react with Boc-amino protected amino acids glycine, β-alanine, and L-lysine activated with 1,1'-carbonyldiimidazole and subsequent dextran ester deprotection.