Every entry summarises what the study set out to test, what it found and why it changes how we design delivery systems. Browse research and reviews or search the collection.
Sérgio Scalzo, Anderson K. Santos, Heloisa A. S. Ferreira, Pedro A. Costa, Pedro H. D. M. Prazeres, Natalia J. A. Da Silva, Lays C. Guimarães, Mário De Morais E Silva, Marco T. R. Rodrigues Alves, Celso T. R. Viana, Itamar C. G. Jesus, Alice P. Rodrigues, Alexander Birbrair, Anderson O. Lobo, Freder
Cardiomyocytes are hard-to-transfect cells, and gene therapy for cardiovascular disease is limited by insufficient delivery to cardiac tissue, nucleic acid degradation, and safety concerns with viral vectors. A safe, effective non-viral platform for pDNA delivery to cardiomyocytes is needed. LNP4 was the top performer: ~1.3-fold higher GFP fluorescence than the second-best LNP8 and ~10-fold higher than the lowest performer LNP6. - LNP4 achieved >60% transfection efficiency at day 2 and >80% at day 4 in.
Rodgers, T., Muzzio, N., Watson, C., & Romero, G. (2021).
Gene-editing tools are large plasmid constructs that cannot spontaneously enter mammalian cells, and viral vectors are limited by cargo size, immunogenicity, and manufacturing challenges. PBAE nanoparticles are promising nonviral carriers, but unmodified PEG-PDHA nanoparticles release encapsulated plasmid DNA too rapidly for efficient intracellular delivery of large gene-editing cargo such as piggyBac transposon. Encapsulation and release: Optimal polymer/plasmid molar ratio = 0.36. Unmodified PEG-PDHA NPs released essentially all PBCAG within 2 h. Layer-by-layer NPs showed no detectable release within 24 h but transfection was.
Synthetic nanoparticles face physiological barriers, RES clearance, and unintended biological interactions despite PEGylation and ligand grafting. Biological vectors—cell membranes, extracellular vesicles (EVs), and viruses—offer biocompatibility, biodegradability, immune evasion, and natural targeting. This review summarizes BNPs integrating biological vectors with functional agents for targeted delivery in imaging and therapy. RBCM-coated PLGA nanoparticles improved half-life by ~50% vs PEGylated nanoparticles and were detected in circulation after 72 h. - NM-NP-CFZ (neutrophil membrane-coated carfilzomib PLGA NPs) reduced lung metastasis by.
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 LNP-nucleic acid therapeutic, for polyneuropathy caused by transthyretin amyloidosis. - Pfizer-BioNTech and Moderna mRNA-LNP COVID-19 vaccines received FDA emergency use authorization.
Bispecific T-cell engager (BiTE) immunotherapy is a promising cancer treatment, but clinical application is limited by high production costs and short in vivo half-life of BiTE proteins. A gene delivery system that enables sustained in vivo expression of BiTE could overcome these limitations, but existing non-viral vectors have low in vivo transfection efficiency. Metal-organic frameworks (MOFs) have been explored for nucleic acid delivery but. ### MOF Characterization & DNA Loading | Parameter | Result | |---------------|------------| | MOF size (TEM) | ~300 nm length × ~100 nm width | | MOF size (DLS) | 187.8 nm | | Zeta potential (MOF) | +24.7 mV | | Zeta.
Margaret M. Billingsley, Nathan Singh, Pranali Ravikumar, Rui Zhang, Carl H. June, Michael J. Mitchell
CAR T cell therapy currently relies on viral vectors or electroporation for T cell engineering. Viral vectors cause permanent CAR expression and carry safety/manufacturing limitations, while electroporation is cytotoxic and requires specialized equipment. A safer, efficient non-viral mRNA delivery platform for human T cells is needed. Seven LNP formulations enhanced luciferase mRNA delivery to Jurkat cells over lipofectamine; C14–4 was the top performer. - Purified C14–4 LNPs: diameter ~65 nm, mRNA encapsulation 86.3%, pKa 6.505; crude C14–4:.
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 personalized, potentially curative therapies. The comment summarizes recent advances and expectations for the near future. Approved therapies: Spinraza (2016), Exondys 51 (2016), Vyondys 53 (2019), Kymriah (2017), Yescarta (2017), Tescartus, Luxturna (2017), Zolgensma (2019). - First CRISPR disease-modifying efficacy: CTX001 for sickle cell.
Wahane A, Waghmode A, Kapphahn A, Dhur K, Gupta A, Bahal R
Nucleic acid analogs and genome-editing proteins have broad therapeutic potential, but clinical translation is limited by poor cell- and organ-specific delivery, endosomal entrapment, enzymatic degradation, and short bioavailability. The review examines lipid- and polymer-based non-viral vectors as strategies to overcome these barriers. DOTMA liposomes: ~100% plasmid DNA entrapment and 5–100-fold higher transfection than calcium phosphate or DEAE-dextran. - SPLPs: ~70 nm particles with ~70% encapsulation; 100–1,000-fold gene expression in distal tumor.
CAR-T cell therapy relies mainly on retroviral/lentiviral vectors, which carry risks of carcinogenicity and complex manufacturing. PiggyBac transposon systems offer a safer, simpler non-viral alternative for stable transgene expression, but primary T lymphocytes are difficult to transfect efficiently. A polymeric nanomicelle carrier was developed to deliver PiggyBac transposon/transposase plasmids to human T cells. Nanomicelle properties: Blank micelles ~62.7 nm; polyplex at N/P 40 ~134.5 nm and +20.9 mV; complete DNA condensation at N/P 20; DTT released DNA. - Transfection at N/P 40: 24.5% ± 3.3% positive cells; viability 82.3% ±.
Haimanti Mandal, Sameer S. Katiyar, Rajan Swami, Varun Kushwah, Parmeshwar B. Katare, Anand Kumar Meka, Sanjay K. Banerjee, Amirali Popat, Sanyog Jain
Non-viral gene delivery vectors are safer than viral vectors but often suffer from lower transfection efficiency and cytotoxicity. ε-Poly-L-lysine (ε-PLL) is a naturally occurring, FDA GRAS cationic polymer with low molecular weight and potential for low toxicity, but its use as a pDNA nanocarrier for efficient in vivo gene delivery was largely unexplored. Physicochemical properties: ε-PLL/pDNA at N/P 50 formed ~194 nm particles with ~+10.9 mV zeta potential. PLL and SuperFect formed larger particles; SuperFect ~800 nm and ~+50 mV. - In vitro transfection: ε-PLL/pDNA was.
Gene therapy has enormous therapeutic potential, but clinical translation is limited by the delivery problem: nucleic acids are negatively charged, unstable, and must overcome multiple biological barriers to reach their intracellular site of action. Viral vectors are efficient but suffer from immunogenicity, limited cargo capacity, insertional mutagenesis risk, and manufacturing challenges. There is a need for innovative synthetic materials that. Ionizable lipid nanoparticles: The most effective formulations for siRNA delivery included lipids with three or more hydrophobic tails, secondary and tertiary amines, and a particle surface pKa of ~5.5–7.0, enabling.
Genada Sinani, Melike Sessevmez, M. Koray Gök, Saadet Özgümüş, Alper Okyar, H. Oya Alpar, Erdal Cevher
Mucosal vaccination can stimulate both mucosal and systemic immunity, but free protein antigens generally induce poor systemic immune responses and require adjuvants. There is a need for a safe, effective nasal vaccine carrier that improves antigen stability, uptake, and immunogenicity, especially for poorly immunogenic protein antigens. Particle properties: RG503 formulation: 234.77 ± 4.49 nm, PDI 0.15, zeta +13.8 ± 4.09 mV, EE 91.16 ± 2.92%. RG502 formulation: EE 84.60 ± 0.67%, zeta +20.1 ± 4.8 mV. Stable for 24 weeks at +4 °C. - Cytotoxicity:.
Jiménez Blanco Jl, Benito Jm, Ortiz Mellet C, García Fernández Jm.
Nonviral gene delivery lacks molecularly well-defined carriers suitable for rigorous structure–activity relationship (SAR) studies. Viral vectors raise immunogenicity, cost, and cargo-size concerns, while polymers, dendrimers, and cationic lipids are often polydisperse or supramolecularly ill-defined. The review argues that “molecular nanoparticles” (MNPs)—monodisperse, shape-persistent molecular entities—can provide programmable, single-isomer. Optimized polycationic amphiphilic cyclodextrins (e.g., 5 and 6) formed 80–100 nm CDplexes at N/P >2 and mediated transfection comparable to or better than PEI/Lipofectamine LTX in COS-7, HeLa, HepG2, and BNL-CL2 cells.
Jing Luo, Caixia Li, Jianlin Chen, Gang Wang, Rong Gao, Zhongwei Gu
Nonviral gene carriers (cationic polymers and lipids) are promising alternatives to viral vectors, but their transfection efficiencies are much lower. While extensive efforts have focused on chemical modification of gene delivery materials, simple optimization of the transfection procedure itself—beyond standard commercial protocols—remains underexplored. A method that can enhance transfection efficiency across multiple cell types without. ### Volume Optimization (PEI, Hep G2, 6-well) | Volume | Transfection Efficiency | vs. Standard | |------------|----------------------------|------------------| | Standard (500 μL complex + 2 mL medium) | Baseline |.
Antonella Mangraviti, Stephany Yi Tzeng, Kristen Lynn Kozielski, Yuan Wang, Yike Jin, David Gullotti, Mariangela Pedone, Nitsa Buaron, Ann Liu, David R. Wilson, Sarah K. Hansen, Fausto J. Rodriguez, Guo-Dong Gao, Francesco Dimeco, Henry Brem, Alessandro Olivi, Betty Tyler, Jordan J. Green
Glioblastoma remains highly lethal with poor prognosis despite surgery, radiation, and chemotherapy. Gene therapy is a promising alternative, but viral vectors carry safety risks. Non-viral polymeric nanoparticles are safer but have historically shown poor in vivo efficacy. A biodegradable poly(beta-amino ester) (PBAE) nanoparticle system was developed to deliver a suicide gene to malignant glioma and improve survival. In vitro transfection: Ten PBAE nanoparticle formulations achieved >50% transfection with <20% toxicity in 9L cells; eight formulations in F98 cells. Three formulations were superior to Lipofectamine 2000 in 9L; fifteen.
Cancer gene therapy requires safe and efficient delivery of nucleic acids, but viral vectors carry risks of immunogenicity and insertional tumorigenicity, while non-viral polymeric vectors are safer but less efficient. The review addresses the need for targeted polymeric nanoparticles that achieve tumor-specific accumulation and cancer-specific transfection through passive targeting, active ligand targeting, and. Representative quantitative findings highlighted in the review: - Transferrin targeting: Tf-conjugated PEG-PEI nanoparticles transfected distant tumor cells in vivo 10–100 times greater than non-tumor cells following.
Gene-based therapy has potential to treat a wide range of diseases, but clinical success is limited by delivery barriers. Viral vectors, while efficient, carry risks of carcinogenesis, immunogenicity, broad tropism, limited packaging capacity, and manufacturing difficulties. Non-viral vectors offer improved safety, larger payload capacity, and easier synthesis, but have low delivery efficiency relative to viruses. Advances in material sciences. ALN-PCS02 (LNP-siRNA targeting PCSK9): Phase I trial showed substantial reduction of PCSK9 and LDL cholesterol with no serious adverse effects. - ALN-TTR02 (Patisiran, LNP-siRNA targeting TTR): Phase II/III; DLinDMA.
Non-viral gene delivery systems are safer than viral vectors but are highly inefficient at nuclear delivery. Only ~10–20% of applied plasmid DNA enters targeted cells, and only ~1–5% of the applied dose reaches the nucleus. The nuclear envelope is a major barrier to transgene delivery, and there is a need for “smart” non-viral vectors that can overcome multiple extracellular and intracellular barriers—especially nuclear transport—to enhance gene. Baseline inefficiency of non-viral vectors: Only 10–20% of applied plasmid DNA enters targeted cells, and only 1–5% enters the nucleus. - NLS enhancement: Addition of NLSs increased luciferase reporter gene expression.
Nupura S. Bhise, Ron B. Shmueli, Jose Gonzalez, Jordan J. Green
Non-viral polymeric nanoparticles are promising for gene therapy and stem cell reprogramming, but the number of plasmids complexed per particle affects transfection efficiency and co-delivery of multiple plasmids. Existing methods for quantifying plasmids per particle have significant drawbacks, and few studies have characterized polymeric nanoparticles in physiologically relevant aqueous conditions. PEI nanoparticles: 90 ± 10 plasmids/particle by NTA; theoretical maximum 267 plasmids/particle. - PBAE nanoparticles: 30 ± 2 to 120 ± 20 plasmids/particle; theoretical maximum 80–195. - Formulation-specific values: -.
Suspension lymphoma/leukemia cells are notoriously hard to transfect with non-viral vectors. Existing methods such as Lipofectamine give poor plasmid DNA delivery in these cells, and viral methods raise safety concerns. A simple, efficient, biodegradable cationic polymer nanocomplex is needed for gene delivery to hard-to-transfect hematopoietic cancer cells. Nanocomplex properties: ~200 nm at pH 7.4, stable for 12 h. At pH <7, nanocomplexes rapidly dissociated and enlarged, consistent with proton-sponge–type release. - Transfection without polybrene: PBAE >7 kDa nanocomplex.
International Journal of Nanomedicine (Dove Press open-access article; exact citation details not fully provided in the 2011ResearchNon-viral Gene Delivery
Stephany Y. Tzeng, Peter H. Yang, Warren L. Grayson, Jordan J. Green
Human umbilical vein endothelial cells (HUVECs) are relevant to tissue engineering, cancer therapy, and angiogenesis-related diseases. Non-viral vectors are safer than viruses but often less efficient. Biodegradable poly(ester amine) (PEA) and poly(amido amine) (PAA) nanoparticles were developed and screened for efficient siRNA and DNA delivery to HUVECs, and to determine whether small changes in polymer structure can tune efficacy for different. siRNA delivery: Up to 60–75% GFP knockdown in HUVECs. Best formulation 454 at 100 w/w achieved 72.3 ± 0.6% knockdown, superior to Lipofectamine 2000 (51.6 ± 5.2%) with comparable or better viability. - DNA delivery: Up.
Nonviral vectors have rarely been tested in neonatal mice because of poor delivery efficiency, yet early gene therapy could prevent irreversible disease, requires lower vector doses due to small body size, and may avoid immune reactions before immune maturation. Viral vectors carry risks such as insertional mutagenesis and immunogenicity, so safe nonviral systemic delivery methods for neonates are needed. Polyplex size: ~143 ± 11 nm. - Optimization: Naked pDNA gave no detectable expression at 24 h. N/P 10 caused mortality (17/21 surviving), N/P 4 gave low expression, and N/P 8 gave strongest expression with 93% survival.
Cationic liposomes (CLs) are attractive non-viral gene-delivery carriers because they are safer, less immunogenic, and easier to prepare than viral vectors, but their transfection efficiency (TE) remains low and the relationship between lipoplex physicochemical properties, lipoplex–cell interaction, and TE is poorly understood. This review describes recent work on CL–DNA complex properties and factors influencing TE, and introduces recent. CL–DNA complexes exhibit structural polymorphism: lamellar \(L_\alpha^C\) and inverted hexagonal \(H_{II}^C\) phases; DOPE tends to adopt the \(H_{II}\) phase and may aid endosomolysis, while cholesterol changes.
Kai K. Ewert, Alexandra Zidovska, Ayesha Ahmad, Nathan F. Bouxsein, Heather M. Evans, Christopher S. Mcallister, Charles E. Samuel, Cyrus R. Safinya
Cationic lipid–nucleic acid (CL-NA) complexes are promising non-viral vectors for gene delivery and siRNA-mediated silencing, but their transfection and silencing efficiencies remain low compared with viral vectors. A major gap is poor mechanistic understanding of how complex structure, membrane charge density, and lipid composition govern cellular pathways and biological activity. For lamellar CL-DNA complexes, TE follows a universal bell-shaped curve as a function of membrane charge density, with an optimal σ_M ≈ 17.0 × 10⁻³ e/Ų. Data for monovalent and multivalent lipids collapse onto this.
