Annals of allergy, asthma & immunology : official publication of the American College of Allergy, Asthma, & Immunology2026ReviewNon-viral Gene Delivery
Schauberger Em, Phillips Ja
The management of allergic and immunologic diseases has long centered on symptom management rather than disease modification. Conventional pharmacotherapy controls manifestations of the immune dysregulation but does little to treat the underlying cause. Newer monoclonal and small molecule therapies target downstream…
Read the article →STAR protocols2026ResearchNon-viral Gene Delivery
Duke L, Kantor B
Adeno-associated vector is a platform of choice for delivering genome-editing toolkits, due to its transient and robust expression. Here, we present a protocol for viral production for in vivo use. We describe steps for transfecting producer cells, harvesting virus, and purifying/concentrating viral particles. We…
Read the article →STAR protocols2026ResearchNon-viral Gene Delivery
Dobner J, Ramachandran H, Rossi A
Genome editing experiments often produce heterogeneous allele mixtures that require structured genotyping. Here, we present a protocol for analyzing amplicon sequencing data from nuclease, base, prime, and mitochondrial base editing experiments using CleanFinder. We describe steps for preparing reference sequences…
Read the article →Molecular therapy. Nucleic acids2026ResearchNon-viral Gene DeliveryDrug Delivery
Olsen Al, Brandt Cbv, Larsen Rk, Fonager Sv, Zubia-Aranburu J, Haskó J, Et Al.
Efficient and cell-type-specific delivery remains a major barrier to CRISPR-based genome editing in vascular endothelial cells. Here, we systematically evaluate lipid nanoparticle (LNP)-mediated mRNA and CRISPR delivery in human umbilical vein endothelial cells (HUVECs). While a conventional SM-102-based LNP…
Read the article →Biomedicines2026ReviewNon-viral Gene DeliveryDrug Delivery
Duffy A, Schweikert A, Tan Sy, Murphy C, Dowling Cm, Hurley K, Et Al.
Chronic lung diseases arise from complex interactions between environmental exposures and host genetic factors. However, these interactions are not fully understood, and despite decades of study, effective curative therapies remain unavailable for many patients. Recent breakthroughs in therapeutic gene-editing have…
Read the article →Theranostics2026ReviewNon-viral Gene DeliveryDrug Delivery
Wu X, Kou X, Bai L, Liu C, Wu Q, Zhang M, Et Al.
Clustered regularly interspaced short palindromic repeats (CRISPR), a precise genome-editing platform, has revolutionized genetic engineering. The highly selective and programmable characteristics of CRISPR have enabled its broad application. Recent advances in CRISPR biotechnology have promoted its widespread use in…
Read the article →Advanced Healthcare Materials2022ReviewNon-viral Gene Delivery
Erin W. Kavanagh And Jordan J. Green
Viral vectors dominate gene therapy but suffer from immunogenicity, insertional mutagenesis risk, cargo-size limits, and manufacturing challenges. Non-viral nanoparticles are safer, more economical, and easier to scale, but their clinical translation has been limited by inefficient delivery, especially for DNA and…
Read the article →2022ReviewNon-viral Gene Delivery
Maeki M, Uno S, Niwa A, Okada Y, Tokeshi M
RNA-loaded lipid nanoparticles (LNPs) are clinically validated for siRNA and mRNA delivery, but their therapeutic performance depends critically on reproducible control of size, size distribution, zeta potential, and RNA encapsulation efficiency. Microfluidic devices offer precise mixing, rapid ethanol dilution, high…
Read the article →Nanomaterials2021ResearchNon-viral Gene Delivery
Tresa Lopez-Royo, Victor Sebastian, Laura Moreno-Martinez, Laura Uson, Cristina Yus, Teresa Alejo, Pilar Zaragoza, Rosario Osta, Manuel Arruebo, Raquel Manzano
CRISPR/Cas gene-editing components are often encoded on large plasmids (9–19 kb), which are difficult to encapsulate and transfect. Most PLGA nanoparticle (NP) gene-delivery studies use small nucleic acids (siRNA) or small plasmids (<6 kb), and the structural integrity and functional performance of large plasmids in…
Read the article →2021ReviewDrug Delivery
Cancer immunotherapy has shown efficacy but is limited by serious adverse effects, nonspecific inflammation, and challenges in spatiotemporal control of immune responses. Nanomedicine and functionalised nanosystems offer programmable pharmacokinetics, co-delivery of immunomodulators, and targeted delivery…
Read the article →Nature Communications2021ReviewNon-viral Gene Delivery
Han X, Zhang H, Butowska K, Swingle Kl, Alameh M-G, Weissman D, Mitchell Mj
RNA therapeutics—ASOs, siRNAs, miRNAs, mRNAs, and CRISPR-Cas9 sgRNAs—are limited by nuclease degradation, large size, and negative charge. Ionisable lipids are the key LNP component that condenses RNA, enables endosomal escape, and reduces toxicity. Since 2008, many ionisable lipids have been created, but the field…
Read the article →Trends in Molecular Medicine2021ReviewNon-viral Gene Delivery
Likely Citation: Swingle Kl, Hamilton Ag, Mitchell Mj
mRNA is degraded by nucleases and cannot easily cross cell membranes because of its large size and negative charge. Delivery therefore requires encapsulation in vehicles such as lipid nanoparticles (LNPs) to enable protein replacement, vaccines, and gene-editing applications. Onpattro was the first FDA-approved…
Read the article →Nature Reviews Materials2021ReviewNon-viral Gene Delivery
Hou X, Zaks T, Langer R, Dong Y
mRNA has therapeutic potential for vaccines, protein replacement, cancer immunotherapy, cellular reprogramming, and genome editing, but it requires safe, effective, and stable delivery systems to protect it from degradation and enable cellular uptake and mRNA release. Lipid nanoparticles have entered the clinic for…
Read the article →Pharmaceutics2021ReviewNon-viral Gene Delivery
Zak Mm, Zangi L
mRNA therapeutics are limited by innate immune activation, rapid RNase degradation, and inefficient delivery to target organs. Although LNPs are the only clinically approved RNA therapeutic carriers, most systemically delivered LNPs accumulate in the liver, so organ-specific delivery remains a major barrier for…
Read the article →Journal of Drug Delivery Science and Technology2021ReviewNon-viral Gene Delivery
Ashok B, Peppas Na, Wechsler Me
CRISPR/Cas9 genome editing holds major therapeutic promise, but in vivo delivery is arguably the most difficult barrier to clinical translation. Efficient, selective transport of editing components to target cells/tissues while minimising off-target effects remains unresolved. This review critiques lipid- and…
Read the article →Nature Biotechnology2020ResearchNon-viral Gene Delivery
David N. Nguyen, Theodore L. Roth, P. Jonathan Li, Peixin Amy Chen, Ryan Apathy, Murad R. Mamedov, Linda T. Vo, Victoria R. Tobin, Daniel Goodman, Eric Shifrut, Jeffrey A. Bluestone, Jennifer M. Puck, Francis C. Szoka, Alexander Marson
CRISPR-Cas9 genome editing in clinically relevant primary cells is limited by inefficient homology-directed repair (HDR) and cytotoxicity of exogenous DNA. Improvements are needed to increase editing efficiency, cell viability, and generalizability across cell types for adoptive cell therapies and research. tCTS…
Read the article →Molecular Therapy: Nucleic Acids2020ResearchNon-viral Gene Delivery
Rui Yan Yuan, Mahita Varanasi, Shanelle Mendes, Hannah M. Yamagata, David R. Wilson, Jordan J. Green
CRISPR-Cas9 gene editing requires efficient intracellular delivery of large cargo. Viral vectors have packaging and production limitations, and nonviral CRISPR plasmid delivery had mainly been studied for single-site knockout, not for gene deletion requiring two cleavage events. The transfection requirements for…
Read the article →Nature Communications2020ReviewNon-viral Gene Delivery
Karen Bulaklak And Charles A. Gersbach
Gene therapy is at an inflection point: recent approvals and clinical successes have validated the field, but broader application is still limited by immune responses to vectors, inefficient delivery outside the retina and liver, and regulatory frameworks that are poorly suited to personalised, potentially curative…
Read the article →Advanced Drug Delivery Reviews2020ReviewDrug Delivery
Nina Filipczak, Jiayi Pan, Satya Siva Kishan Yalamarty, Vladimir P. Torchilin
Liposomes remain an important nano-sized drug delivery system because of their biomimetic bilayer, ease of preparation, and biocompatibility, but clinical translation is still limited by reproducibility, scale-up, stability, and payload-specific barriers. The review updates recent advances in liposome preparation…
Read the article →Cancer Letters2019ReviewNon-viral Gene Delivery
Mukalel Aj, Riley Rs, Zhang R, Mitchell Mj.
Nucleic acid therapeutics—DNA, mRNA, and CRISPR/Cas9 genome editing systems—hold significant potential for cancer immunotherapy but face major delivery barriers, including rapid in vivo degradation, poor cellular uptake, required nuclear entry, and toxicity in healthy tissues. Nanoparticle delivery systems are being…
Read the article →Biomaterials2019ReviewNon-viral Gene Delivery
Zhao W, Hou X, Vick Og, Dong Y.
Genetic and rare diseases (GARDs) affect more than 350 million patients worldwide, and many lack effective treatments. RNA-based therapeutics—siRNA, mRNA, miRNA, lncRNA, and RNA-containing genome editing systems—are promising, but naked RNA has poor cellular penetration, is degraded in biological fluids, and does not…
Read the article →Nature Communications2018ResearchNon-viral Gene Delivery
Ewoud B. Compeer, Felix Kraus, Manuela Ecker, Gregory Redpath, Mayan Amiezer, Nils Rother, Philip R. Nicovich, Natasha Kapoor-Kaushik, Qiji Deng, Guerric P.B. Samson, Zhengmin Yang, Jieqiong Lou, Michael Carnell, Haig Vartoukan, Katharina Gaus, Jérémie Rossy
Polarized endocytic recycling of the T cell receptor (TCR) to the immunological synapse is essential for T cell activation, yet the cellular mechanisms that coordinate internalisation of surface receptors with sustained delivery back to the plasma membrane remain incompletely understood. The role of flotillins in…
Read the article →Small Methods.2018ReviewNon-viral Gene Delivery
Evers Mjw, Kulkarni Ja, Van Der Meel R, Cullis Pr, Vader P, Schiffelers Rm
Free siRNA, mRNA, pDNA, and CRISPR/Cas9 components are rapidly degraded and cleared, so delivery systems are required. LNPs are the most clinically advanced non-viral siRNA carriers; Patisiran reached Phase III and market application in late 2017. The field needed a clearer understanding of LNP design parameters and…
Read the article →Frontiers in Pharmacology2018ReviewDrug Delivery
Sima Rezvantalab, Natascha Ingrid Drude, Mostafa Keshavarz Moraveji, Nihan Güvener, Emily Kate Koons, Yang Shi, Twan Lammers, Fabian Kiessling
PLGA-based nanoparticles are widely investigated for cancer therapy, but their clinical translation remains limited. A consolidated overview was needed of how PLGA NP physicochemical properties, biological behaviour, targeting strategies, and combination therapies interact—and why promising preclinical results have…
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