Aug 2026· 3 Biotech· Vol 16· 0 citations· 224 references
Medicine
TL;DR
This review provides a comprehensive overview of the current status of viral and non-viral vector systems for in vivo and ex vivo applications, and key comparisons are made across safety, efficacy, scalability, and immune responses.
Abstract
Gene and cell therapies have emerged as transformative approaches for treating a wide range of genetic and acquired diseases. Central to their success is the development of safe and effective gene delivery systems, categorized broadly into viral and non-viral vectors. Each system has its own advantages and limitations, requiring careful consideration of the target tissue, disease, and the balance between safety, efficacy, and scalability. Viral vectors, including adeno-associated viruses, retroviruses, lentiviruses, herpes simplex viruses, and adenoviruses, offer high transduction efficiency and specificity but raise concerns about immunogenicity and production challenges. Non-viral systems, such as lipid nanoparticles (LNPs) and other synthetic carriers, provide scalable, cost-effective, and potentially safer alternatives but often face hurdles in transduction efficiency and targeted delivery. This review provides a comprehensive overview of the current status of these vector systems for in vivo and ex vivo applications. Key comparisons are made across safety, efficacy, scalability, and immune responses, highlighting recent advancements and innovative approaches. We also discuss the outlook for next-generation gene transfer technologies, focusing on improvements in vector design, manufacturing, and application versatility. By addressing these considerations, we aim to inform the development of optimized therapeutic strategies that leverage the unique strengths of each delivery system.
Lentiviral vectors have revolutionized gene therapy by efficient and stable transduction of dividing and non-dividing cells, their large packaging capacity, and their compatibility with pseudotyping to alter viral tropism. The vesicular stomatitis virus glycoprotein (VSV-G) is widely used as a viral envelope protein of choice to pseudotype lentiviral vector particles as it confers exceptional particle stability and a broad tropism, due to the ubiquitous nature of the low-density lipoprotein receptor (LDLR). While this broad tropism facilitates transduction of diverse cell types, it precludes accurate in vivo targeting of specific cell populations. Structural insights into VSV-G have made receptor-blinding possible and revealed sites amenable to mutation while preserving fusion capacity. Coupled with targeting moieties, VSV-G pseudotyped lentiviral particles are redirected towards cells expressing target antigens. Such targeted vectors open new possibilities for in vivo gene therapy across oncology, infectious diseases, transplantation medicine, and other diseases. Use of targeted vectors will make in vivo gene therapy more accessible than cost-intensive ex vivo gene therapies. Since targeted vectors will be available as 'off-the-shelf' drugs, they will also drastically reduce time-to-treatment. This review highlights advances in bioengineering to exploit the versatility of VSV-G-pseudotyped lentiviral vectors and explores their vast potential for targeted gene delivery.
Anjali Shrivastava, Felix L. Warnecke, J. Schott et al.· Molecular Therapy· 0 citations
Abstract Gene therapy has emerged as a transformative strategy for treating a wide range of genetic and acquired disorders. Despite its potential, clinical translation is hindered by the limitations of viral vectors, including immunogenicity, insertional mutagenesis, limited cargo capacity, and production challenges. Consequently, non-viral gene delivery systems have gained increasing attention as safer, more versatile alternatives. These platforms, including lipid-based nanoparticles, polymers, dendrimers, inorganic nanocarriers, and hybrid systems, offer customizable physicochemical properties, scalable manufacturing, and reduced risk of adverse immune responses. This review systematically expresses recent advances in non-viral vectors, focusing on the key parameters that influence cellular uptake, endosomal escape, nuclear localization, and overall transfection efficiency in various disease, especially cancers. Additionally, we evaluate recent preclinical and clinical studies, highlighting promising translational outcomes and therapeutic applications. By comparing different non-viral strategies and discussing their mechanistic underpinnings, this review underscores the potential of non-viral vectors to overcome the inherent limitations of viral delivery and to drive the development of next-generation gene therapy approaches that are safer, more adaptable, and clinically relevant.
Fatemeh Madani, Elaheh Izadi, Maral Motamedi et al.· International Journal of Nan...· 0 citations
This article aims to provide a working framework for verifying the potency, genomic integrity, and clinical safety of vector-based gene therapies—one intended to be useful both to laboratories developing these products and to those responsible for regulating them.
Yusra A. Radeef, Z. Abdullah, Eman Fadhel Abbas Awadh· International Journal of Mul...· 0 citations
Helper-dependent adenoviral vectors (HDAdVs), which lack all viral coding sequences, enable the delivery of up to 35 kilobases (kb) of therapeutic DNA into target cells. This large packaging capacity facilitates the transfer of complex, large, or even multiple transgenes. Despite the technical complexity of their production, HDAdVs have been extensively evaluated in numerous preclinical studies, and several clinical trials have recently been initiated. This review provides a comprehensive, state-of-the-art overview of recent advances in the HDAdVs technology. It summarizes key strategies for vector development, including capsid engineering and improvements in production methodologies. Furthermore, it reviews preclinical in vivo studies with a focus on vector design, target indications, and therapeutic outcomes. Most preclinical applications have focused on targeting the liver and hematopoietic stem cells; however, additional organ systems and vaccination approaches based on HDAdVs are also being actively explored. In addition to preclinical progress, three recently initiated clinical trials using HDAdVs targeting knee osteoarthritis, chronic granulomatous disease, and solid tumors are highlighted. Nevertheless, several challenges must be addressed to enable broader clinical translation of HDAdVs. These include the complexity of large-scale manufacturing, achieving efficient and specific vector targeting, and overcoming host immune responses. In conclusion, HDAdV-based vector systems hold considerable promise for the treatment of a wide range of diseases. Continued technological advancements, including the development of next-generation adenoviral platforms and the integration of precision gene therapy approaches, are likely to further enhance their potential as versatile biotherapeutic tools.
Montaha Alshawabkeh, Eric Ehrke-Schulz, Wenli Zhang et al.· Human Gene Therapy· 0 citations
Content of image described in text. Abstract Background Gene therapy has emerged as a transformative approach for treating diseases that are caused by genetic defects, including cancer, inherited metabolic disorders and immunodeficiencies. However, its clinical success depends critically on the availability of safe, efficient and non-immunogenic delivery systems. Conventional viral vectors, while highly effective, are associated with immunogenicity, oncogenic risk and high production costs. Non-viral alternatives offer improved safety but they have lower transfection efficiency and limited stability. Exosomes are naturally secreted, endosome-derived nanovesicles ranging from 30–150 nm in diameter and have emerged as promising next-generation carriers for gene and drug delivery. Their inherent biocompatibility, low immunogenicity, ability to cross biological barriers and capacity to transport diverse molecular cargo including nucleic acids, proteins and small molecules make them particularly attractive as therapeutic platforms. Methods This review summarizes the biogenesis, molecular composition and classification of exosomes and examines their applications in siRNA/miRNA delivery, CRISPR/Cas9 gene editing, mRNA therapy, cancer treatment and vaccine development. Special attention is given to engineered, plant-derived and stem cell-derived exosomes, as well as current cargo-loading strategies and commercial therapeutic platforms. Results Despite their promise, significant challenges remain, including low cargo-loading efficiency, batch heterogeneity, limited scalability and the absence of standardized manufacturing and regulatory frameworks. Conclusions Future research must address these barriers to accelerate the clinical translation of exosome-based therapeutics.
Elza Karabagh, Babek Alibayov, Adil Allahverdiyev· Expert Reviews in Molecular...· 0 citations
Objective: To review the existing genetic engineering strategies commonly used for generating chimeric antigen receptor (CAR)-T cell therapy. Methods: This article provides a comprehensive literature review and compares different approaches on viral, non-viral and precision genome editing technologies, aiming to provide an overview and guidelines for methods selection during clinical CAR-T cell manufacturing. Results: CAR-T cell therapy has revolutionized cancer treatment for hematological malignancies. The manufacturing of CAR-T cell therapy relies on the genetic modification of T cells to achieve ectopic CAR gene expression. Depending on the evolving CAR designs, viral, non-viral, and genome editing platforms are used for optimal CAR expression, which could determine the clinical efficacy and safety profiles of these products. Efforts have been ongoing to empower CAR-T cell efficacy while minimizing toxicity over the past decades. Conclusion: Successful CAR-T cell therapy depends on rational platform selection and optimization of gene delivery methods based on clinical needs and context.
Wing Keung Chan, M. Lima· JOURNAL OF BONE MARROW TRANS...· 0 citations