Biomaterials Science2022ReviewNon-viral Gene Delivery
Sydney R. Shannon, Elena Ben-Akiva, Jordan J. Green
Gene therapies are transforming treatment for many diseases, but clinical efficacy and safety depend on both the delivery material and the cargo. Non-viral delivery to immune cells remains especially challenging, and no clinical translation breakthrough has yet been achieved for non-viral gene editing. This mini-review addresses that gap by surveying biomaterial-based delivery to immune cells, CRISPR/Cas9 cargo options, and how the two fields can be integrated for cancer immunotherapy. --- - PBAE nanoparticles with anti-CD3ε Fab2, MTAS, and NLS delivered DNA encoding leukemia-specific CARs to T cells at 34% efficiency *in vivo*, programming functional antigen recognition and anti-tumor effects. - PBAE mRNA CAR nanoparticles (anti-CD8, PGA-coated) transiently transfected T cells at 10% efficiency *in vivo*. - CART polymers: ~80% mRNA transfection in Jurkat T cells *in vitro*, but only ~1.5% *in vivo*; mixed hydrophobic blocks outperforme
Read the article →Signal Transduction and Targeted Therapy2022ReviewNon-viral Gene Delivery
Qin S, Tang X, Chen Y, Et Al.
mRNA therapeutics have fueled hope to combat incurable diseases, but insufficient understanding of mRNA instability, immunogenicity, and delivery has impeded progress. The review argues that mRNA-based drugs—especially after the COVID-19 vaccine success—can become powerful, versatile tools, and that mRNA optimization and delivery systems are the key bottlenecks to solve. COVID-19 mRNA vaccines: ~90% effectiveness for full vaccination, 80% for partial; BNT162b2 95% efficacy in phase III; mRNA-1273 94.1% efficacy; Omicron-neutralizing antibodies largely undetectable in most recipients,.
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 gene-editing cargoes that must reach the nucleus. There is a need to review non-viral nanomaterial design, delivery barriers, and. Viral vector dominance: Approximately 70% of gene therapy clinical trials to date have used viruses. - LNP gene editing in liver: A single administration of CRISPR-Cas9 LNPs achieved >97% reduction in target protein.
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 reproducibility, high-throughput formulation screening, and continuous production, making them attractive for standardized LNP. Increasing PEG-lipid from 1 to 5 mol% reduced siRNA-LNP size from 54 nm to 28 nm; DLin-KC2-DMA LNPs achieved 50% FVII silencing at 0.01 mg/kg in mice. - Optimized mRNA-LNP formulations improved delivery and expression.
Read the article →Stem Cell Research & Therapy2021ReviewNon-viral Gene Delivery
Farough Marofi, Roza Motavalli, Vladimir A. Safonov, Lakshmi Thangavelu, Alexei Valerievich Yumashev, Markov Alexander, Navid Shomali, Max Stanley Chartrand, Yashwant Pathak, Mostafa Jarahian, Sepideh Izadi, Ali Hassanzadeh, Naghmeh Shirafkan, Safa Tahmasebi, Farhad Motavalli Khiavi
CAR T cell therapy has transformed treatment of hematologic malignancies, but its efficacy in solid tumors remains unsupported. Solid tumors present distinct barriers—antigen heterogeneity, poor T cell trafficking/infiltration, and an immunosuppressive tumor microenvironment (TME)—that limit CAR T cell function. This review addresses these barriers and evaluates emerging strategies to improve CAR T cell therapy in non-hematologic malignancies. --- - Ovarian cancer: MSLN-CAR NK cells significantly killed MSLN⁺ ovarian cancer cells (SK-OV-3, OVCAR-3) *in vitro*. MUC16-specific CAR T cells eradicated malignant cells in mouse models. TAG72-CAR T and FRα-CAR T inhibited ovarian cancer growth. - Breast cancer: MUC28z CAR T cells (targeting tMUC1) reduced TNBC tumor proliferation and survival in a xenograft model. HRG1β-based CAR T cells inhibited breast cancer via HER family receptors. Anti-HER2 CAR T cells triggered cell death in HER2-overex
Read the article →Blood Cancer Journal2021ReviewNon-viral Gene Delivery
Robert C. Sterner, Rosalie M. Sterner
CAR-T cell therapy has produced remarkable clinical responses in certain B cell leukemias and lymphomas, but major limitations remain—including life-threatening toxicities, limited efficacy in solid tumors, antigen escape, poor persistence/trafficking, and an immunosuppressive microenvironment. This review addresses these barriers and discusses recent innovations in CAR-T engineering to improve efficacy and safety in both hematological malignancies and solid tumors. --- - Antigen escape: 70–90% of relapsed/refractory ALL patients show durable responses to CD19 CAR-T, but 30–70% of recurrent disease involves CD19 downregulation/loss. BCMA loss also observed in multiple myeloma. - Dual targeting: CD19/CD22 and CD19/BCMA dual-targeted CAR-T cells show promising efficacy and favorable safety in early clinical trials. Tandem HER2/IL13Rα2 CARs improved anti-tumor activity and decreased antigen escape in glioblastoma models. - Toxicity reducti
Read the article →EBioMedicine* (Elsevier)2021ReviewNon-viral Gene Delivery
Ella S. Atsavapranee, Margaret M. Billingsley, Michael J. Mitchell
Adoptive T cell therapy has been revolutionized by genetic engineering, but viral transduction—the standard delivery method—has limitations including small cargo capacity, high immunogenicity, high manufacturing cost, and risks of genotoxicity/insertional mutagenesis. There is a need for improved non-viral delivery technologies to enable precise gene editing in T cells for cancer immunotherapy, especially for solid tumours and allogeneic applications. --- - Five FDA-approved CAR T cell therapies: Kymriah, Yescarta, Tecartus, Breyanzi, and Abecma. - Sleeping Beauty clinical trials: Patients with advanced NHL and ALL undergoing HSCT and CAR T cell infusion showed no acute or latent toxicities and no exacerbation of GVHD. - CRISPR/Cas9 first-in-human trial: T cells from three patients with refractory cancer were edited to remove endogenous TCRs and PD-1, achieving highly specific editing at targeted loci without clinical toxicity; edited T
Read the article →Journal of Materials Chemistry B2021ReviewNon-viral Gene Delivery
Rih-Yang Huang, Zhuo-Hao Liu, Wei-Han Weng, Chien-Wen Chang
Gene therapy requires safe, precise, efficient, and cost-effective delivery. Viral vectors are efficient but face manufacturing, cost, and safety challenges; non-viral vectors are safer and more scalable but often have low delivery efficiency. Magnetic nanoparticles (MNPs) have emerged as a promising strategy to enhance viral and non-viral gene delivery under an external magnetic field, while also enabling magnetic targeting, MRI tracking, and magnetic hyperthermia. --- - Magnetofection speed: MNP-based gene carriers can be attracted to cell surfaces in a few minutes, compared with hours for regular transfection. - Transfection enhancement: EMF enhanced lipid/SPIO-mediated transfection sixfold. PEI/Si@MNP with Dox and P-gp shRNA showed magnetically targeted delivery under 0.42 T for 12 h. - In vivo neuronal transfection: NeuroMag delivered EYFP-channelrhodopsin to rat visual cortex neurons with 72.66% and 86.63% expression at 3 and 30 d
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 PLGA NPs remain poorly characterized. There is a need to compare synthesis methods for encapsulating large plasmids without. pDNA stability: Batch ultrasound double emulsion completely degraded the 9.4 kb plasmid (standard) or left only 8.80% supercoiled (modified). Microfluidics-assisted double emulsion avoided degradation but produced only.
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 protein replacement, cancer immunotherapy, and gene editing. LNPs are the only RNA therapeutic carriers approved for clinical use at the time of the review. - PEG content from 1% to 5% produces LNPs approximately 100 nm to 20 nm in size; 0.5% PEG gave highest subretinal.
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 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.
Read the article →Molecular Therapy2019ReviewNon-viral Gene Delivery
Piotr S. Kowalski, Arnab Rudra, Lei Miao, Daniel G. Anderson
mRNA has broad therapeutic potential for vaccination, protein replacement, and gene editing, but its clinical translation is limited by the need for improved intracellular delivery systems. mRNA is much larger than siRNA or ASOs (~300–5,000 kDa, ~1–15 kb vs ~14 kDa and 4–10 kDa, respectively), making delivery more challenging. This review addresses the gap between mRNA's therapeutic promise and the delivery barriers that still restrict its broad application. --- - Protein expression half-life: Modified mRNA protein production half-life ranges from ~50 h in vitro to 7–30 h in vivo, depending on route. Circular RNAs provide up to 3-fold increase in half-life in vitro. - Liver expression: LNPs with lipid 5 achieved hEPO expression at doses as low as 0.01 mg/kg in NHPs, with peak expression at 6–12 h and sustained for over a month with weekly dosing. - Gene editing: LNP-mediated co-delivery of Cas9 mRNA and sgRNA achieved >80% indels for PC
Read the article →Biomaterials Science2019ResearchNon-viral Gene Delivery
Brynn R. Olden, Emmeline Cheng, Yilong Cheng, Suzie H. Pun
T cells are an important target for ex vivo gene delivery and editing, but most commercial reagents cannot transfect them and cationic polymers show only moderate success. The specific intracellular barriers limiting non-viral gene delivery to human T cells are poorly understood, hindering rational polymer design. Uptake: Polyplex uptake was significantly lower in primary human T cells than in HeLa or Jurkat cells. VIPER uptake was higher than Comb in Jurkat and HeLa, but uptake was not predictive of transfection in primary T.
Read the article →Nature Reviews Materials2017ReviewNon-viral Gene Delivery
Khalid A. Hajj And Kathryn A. Whitehead
mRNA therapeutics have enormous potential for vaccines, cancer immunotherapy, protein replacement, and gene editing, but widespread clinical application is limited by the lack of safe and effective delivery vehicles. Naked mRNA is large (~10⁵–10⁶ Da), densely negatively charged, and rapidly degraded by nucleases; its cellular uptake rate is less than 1 in 10,000 molecules, and its intracellular half-life is only ~7 hours. There is a critical. Naked mRNA uptake: Cellular uptake rate of naked mRNA is <1 in 10,000 molecules; median intracellular half-life is only ~7 hours. - Endosomal escape bottleneck: Even world-class RNA delivery materials escape the.
Read the article →Molecular Therapy2017ReviewNon-viral Gene Delivery
Cullis Pr, Hope Mj
Genetic drugs such as siRNA, mRNA, and plasmid DNA could treat most diseases by silencing pathological genes, expressing therapeutic proteins, or enabling gene editing. However, naked RNA/DNA is rapidly degraded, does not accumulate in target tissues, and cannot cross cell membranes efficiently. Lipid nanoparticles (LNPs) are the leading non-viral delivery system to enable clinical gene therapies. LNP siRNA systems containing MC3 can silence hepatocyte genes at doses as low as 0.005 mg siRNA/kg in mice. - MC3 improved gene-silencing activity by over three orders of magnitude compared with DLinDMA. - Optimal.
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