The DOI matches its Crossref record; 153 of the 153 references it cites were found in a registry or an index.
State-of-the-art design and rapid-mixing production techniques of lipid nanoparticles for nucleic acid delivery
Open accessAbstract
Abstract Lipid nanoparticles (LNPs) are currently the most clinically advanced nonviral carriers for the delivery of small interfering RNA (siRNA). Free siRNA molecules suffer from unfavorable physicochemical characteristics and rapid clearance mechanisms, hampering the ability to reach the cytoplasm of target cells when administered intravenously. As a result, the therapeutic use of siRNA is crucially dependent on delivery strategies. LNPs can encapsulate siRNA to protect it from degradative endonucleases in the circulation, prevent kidney clearance, and provide a vehicle to deliver siRNA in the cell and induce its subsequent release into the cytoplasm. Here, the structure and composition of LNP–siRNA are described including how these affect their pharmacokinetic parameters and gene‐silencing activity. In addition, the evolution of LNP–siRNA production methods is discussed, as the development of rapid‐mixing platforms for the reproducible and scalable manufacturing has facilitated entry of LNP–siRNA into the clinic over the last decade. Finally, the potential of LNPs in delivering other nucleic acids, such as messenger RNA and CRISPR/Cas9 components, is highlighted alongside how a design‐of‐experiment approach may be used to improve the efficacy of LNP formulations.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“The latter contains positively charged ionizable amine groups (at low pH) to interact with the anionic mRNA during particle formation and also facilitate membrane fusion during internalization (Evers et al., 2018, Reichmuth et al., 2016).”
From mRNA-lipid nanoparticle COVID-19 vaccines: structure and stability (2021) · cites this paper as Evers et al., 2018
In the citing paper
“Together with the mRNA, these components form particles of about 60–100 nm in size by using a rapid mixing production technique (Evers et al., 2018).”
From mRNA-lipid nanoparticle COVID-19 vaccines: structure and stability (2021) · cites this paper as Evers et al., 2018
In the citing paper
“Note that the physicochemical and structural characteristics of mRNA LNPs will depend on the lipid composition, the ratio of mRNA to the total amount of (cationic) lipids, and the LNP synthesis (for a detailed review on these parameters also see [107]).”
From Three decades of messenger RNA vaccine development (2019) · cites this paper as [107]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., & Mello, C. C. (1998) · Nature · doi:10.1038/35888 · Cited by 15,429
Genome engineering using the CRISPR-Cas9 system
Ran, F. A., Hsu, P. D., Wright, J., Agarwala, V., Scott, D. A., & Zhang, F. (2013) · Nature Protocols · doi:10.1038/nprot.2013.143 · Cited by 11,133
The new frontier of genome engineering with CRISPR-Cas9
Doudna, J. A., & Charpentier, E. (2014) · Science · doi:10.1126/science.1258096 · Cited by 7,062
Principles of nanoparticle design for overcoming biological barriers to drug delivery
Blanco, E., Shen, H., & Ferrari, M. (2015) · Nature Biotechnology · doi:10.1038/nbt.3330 · Cited by 7,002
Recent advances with liposomes as pharmaceutical carriers
Torchilin, V. P. (2005) · Nature Reviews Drug Discovery · doi:10.1038/nrd1632 · Cited by 5,282
Direct Gene Transfer into Mouse Muscle in Vivo
Wolff, J. A., Malone, R. W., Williams, P., Chong, W., Acsadi, G., Jani, A., & Felgner, P. L. (1990) · Science · doi:10.1126/science.1690918 · Cited by 4,173
Strategies in the design of nanoparticles for therapeutic applications
Petros, R. A., & DeSimone, J. M. (2010) · Nature Reviews Drug Discovery · doi:10.1038/nrd2591 · Cited by 3,651
Chaotic Mixer for Microchannels
Stroock, A. D., Dertinger, S. K. W., Ajdari, A., Mezić, I., Stone, H. A., & Whitesides, G. M. (2002) · Science · doi:10.1126/science.1066238 · Cited by 3,435
Non-viral vectors for gene-based therapy
Yin, H., Kanasty, R. L., Eltoukhy, A. A., Vegas, A. J., Dorkin, J. R., & Anderson, D. G. (2014) · Nature Reviews Genetics · doi:10.1038/nrg3763 · Cited by 3,123
Knocking down barriers: advances in siRNA delivery
Whitehead, K. A., Langer, R., & Anderson, D. G. (2009) · Nature Reviews Drug Discovery · doi:10.1038/nrd2742 · Cited by 3,115
Research works printed but not linked: 5
- Leung A. K. K., 2015, Ph.D. Thesis
- Mozafari M. R., 2010, Liposomes, Method in Molecular Biology (Methods and Protocols)
- Walsh C., 2014, Drug Delivery Systems, 109
- Hope M., 1993, Liposome Technology, 123
- MacLachlan I., 1999, Curr. Opin. Mol. Ther., 1, 252
Cited by
Sorted by citations
Tenchov, R., Bird, R., Curtze, A. E., & Zhou, Q. (2021) · ACS Nano · doi:10.1021/acsnano.1c04996 · Cited by 2,221
mRNA-lipid nanoparticle COVID-19 vaccines: structure and stability
Schoenmaker, L., Witzigmann, D., Kulkarni, J. A., Verbeke, R., Kersten, G., Jiskoot, W., & Crommelin, D. J. A. (2021) · International Journal of Pharmaceutics · doi:10.1016/j.ijpharm.2021.120586 · Cited by 1,566
“The latter contains positively charged ionizable amine groups (at low pH) to interact with the anionic mRNA during particle formation and also facilitate membrane fusion during internalization (Evers et al., 2018; Reichmuth et al., 2016).”
Self-citationmRNA vaccine for cancer immunotherapy
Miao, L., Zhang, Y., & Huang, L. (2021) · Molecular Cancer · doi:10.1186/s12943-021-01335-5 · Cited by 966
Chemistry of Lipid Nanoparticles for RNA Delivery
Eygeris, Y., Gupta, M., Kim, J., & Sahay, G. (2021) · Accounts of Chemical Research · doi:10.1021/acs.accounts.1c00544 · Cited by 917
Nanomaterial Delivery Systems for mRNA Vaccines
Buschmann, M. D., Carrasco, M. J., Alishetty, S., Paige, M., Alameh, M. G., & Weissman, D. (2021) · Vaccines · doi:10.3390/vaccines9010065 · Cited by 582
“Prior methods, such as thin film hydration and ethanol injection, are generally not used since they result in heterogeneous larger-sized nanoparticles with lower mRNA encapsulation efficiency, which are difficult to scale up [95].”
BackgroundMethodLipid nanoparticles enabling gene therapies: from concepts to clinical utility
Kulkarni, J. A., Cullis, P. R., & van der Meel, R. (2018) · Nucleic Acid Therapeutics · doi:10.1089/nat.2018.0721 · Cited by 592
Wang, X., Liu, S., Sun, Y., Yu, X., Lee, S. M., Cheng, Q., Wei, T., Gong, J., Robinson, J. J., Zhang, D., et al. (2022) · Nature Protocols · doi:10.1038/s41596-022-00755-x · Cited by 581
Microfluidic formulation of nanoparticles for biomedical applications
Shepherd, S. J., Issadore, D., & Mitchell, M. J. (2021) · Biomaterials · doi:10.1016/j.biomaterials.2021.120826 · Cited by 465
Carrasco, M. J., Alishetty, S., Alameh, M., Said, H., Wright, L., Paige, M. A., Soliman, O. Y., Weissman, D., Cleveland, T. E., Grishaev, A., et al. (2021) · Communications Biology · doi:10.1038/s42003-021-02441-2 · Cited by 460
Lipid nanoparticle technology for therapeutic gene regulation in the liver
Witzigmann, D., Kulkarni, J. A., Leung, J., Chen, S., Cullis, P. R., & van der Meel, R. (2020) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2020.06.026 · Cited by 376
A Comprehensive Review of the Global Efforts on COVID-19 Vaccine Development
Li, Y., Tenchov, R., Smoot, J., Liu, C., Watkins, S., & Zhou, Q. A. (2021) · ACS Central Science · doi:10.1021/acscentsci.1c00120 · Cited by 348
Three decades of messenger RNA vaccine development
Verbeke, R., Lentacker, I., De Smedt, S. C., & Dewitte, H. (2019) · Nano Today · doi:10.1016/j.nantod.2019.100766 · Cited by 342
Nucleic acid delivery for therapeutic applications
Gupta, A., Andresen, J. L., Manan, R. S., & Langer, R. (2021) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2021.113834 · Cited by 351
Scalable mRNA and siRNA Lipid Nanoparticle Production Using a Parallelized Microfluidic Device
Shepherd, S. J., Warzecha, C. C., Yadavali, S., El-Mayta, R. D., Alameh, M., Wang, L., Weissman, D., Wilson, J. M., Issadore, D., & Mitchell, M. J. (2021) · Nano Letters · doi:10.1021/acs.nanolett.1c01353 · Cited by 324
Synthetic materials at the forefront of gene delivery
Lostalé‐Seijo, I., & Montenegro, J. (2018) · Nature Reviews Chemistry · doi:10.1038/s41570-018-0039-1 · Cited by 315
Lipid nanoparticles for delivery of RNA therapeutics: Current status and the role of in vivo imaging
Jung, H. N., Lee, S.-Y., Lee, S., Youn, H., & Im, H.-J. (2022) · Theranostics · doi:10.7150/thno.77259 · Cited by 282
“4), the charge of the ionizable lipids becomes less positive or neutral [23, 24].”
BackgroundMethodStealth nanoparticles in oncology: Facing the PEG dilemma
Zalba, S., ten Hagen, T. L. M., Burgui, C., & Garrido, M. J. (2022) · Journal of Controlled Release · doi:10.1016/j.jconrel.2022.09.002 · Cited by 292
Circular RNA vaccine in disease prevention and treatment
Niu, D., Wu, Y., & Lian, J. (2023) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-023-01561-x · Cited by 281
“158 Cholesterol outside LNPs Fig.”
Lipid-based nanoparticle technologies for liver targeting
Böttger, R., Pauli, G., Chao, P., Fayez, N. A., Hohenwarter, L., & Li, S. (2020) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2020.06.017 · Cited by 277
From influenza to COVID-19: Lipid nanoparticle mRNA vaccines at the frontiers of infectious diseases
Pilkington, E. H., Suys, E. J. A., Trevaskis, N. L., Wheatley, A. K., Zukancic, D., Algarni, A., Al-Wassiti, H. A., Davis, T. P., Pouton, C. W., Kent, S. J., et al. (2021) · Acta Biomaterialia · doi:10.1016/j.actbio.2021.06.023 · Cited by 269
“Liposomes, meanwhile, generally exhibit low enapsulation efficiency of oligonucleotides [108] , making them imractical for development towards in vivo transport and delivery of”
Polysarcosine-functionalized lipid nanoparticles for therapeutic mRNA delivery
Nogueira, S. S., Schlegel, A., Maxeiner, K., Weber, B., Barz, M., Schroer, M. A., Blanchet, C. E., Svergun, D. I., Ramishetti, S., Peer, D., et al. (2020) · ACS Applied Nano Materials · doi:10.1021/acsanm.0c01834 · Cited by 252
Webb, C., Forbes, N., Roces, C. B., Anderluzzi, G., Lou, G., Abraham, S., Ingalls, L., Marshall, K., Leaver, T., Watts, J. A., et al. (2020) · International Journal of Pharmaceutics · doi:10.1016/j.ijpharm.2020.119266 · Cited by 206
Lipid nanoparticles for siRNA delivery in cancer treatment
Moukhtari, S. H. E., Garbayo, E., Amundarain, A., Pascual‐Gil, S., Carrasco‐Leon, A., Prósper, F., Agirre, X., & Blanco‐Prieto, M. J. (2023) · Journal of Controlled Release · doi:10.1016/j.jconrel.2023.07.054 · Cited by 221
Wahane, A. V., Waghmode, A., Kapphahn, A., Dhuri, K., Gupta, A., & Bahal, R. (2020) · Molecules · doi:10.3390/molecules25122866 · Cited by 202
“Currently, LNPs are formulated using various methods; T-junction mixing, microfluidic hydrodynamic focusing (MHF), and staggered herringbone mixing (SHM) that contains ethanol as a phase to solubilize the lipids [29].”
BackgroundMethodAnionic Lipid Nanoparticles Preferentially Deliver mRNA to the Hepatic Reticuloendothelial System
Pattipeiluhu, R., Arias‐Alpizar, G., Basha, G., Chan, K. Y. T., Bussmann, J., Sharp, T. H., Moradi, M., Sommerdijk, N., Harris, E. N., Cullis, P. R., et al. (2022) · Advanced Materials · doi:10.1002/adma.202201095 · Cited by 194
CHARMM-GUI Membrane Builder : Past, Current, and Future Developments and Applications
Feng, S., Park, S., Choi, Y. K., & Im, W. (2023) · Journal of Chemical Theory and Computation · doi:10.1021/acs.jctc.2c01246 · Cited by 197
Lipid nanoparticle-mediated drug delivery to the brain
Khare, P., Edgecomb, S. X., Hamadani, C. M., Tanner, E. E. L., & S Manickam, D. (2023) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2023.114861 · Cited by 187
Recent Advances in the Lipid Nanoparticle-Mediated Delivery of mRNA Vaccines
Swetha, K., Kotla, N. G., Tunki, L., Jayaraj, A., Bhargava, S. K., Hu, H., Bonam, S. R., & Kurapati, R. (2023) · Vaccines · doi:10.3390/vaccines11030658 · Cited by 196
“Moreover, the biocompatibility of the LNPs is enhanced as cholesterol is one of the major components of biological membranes [25].”
BackgroundPlatforms for Production of Protein-Based Vaccines: From Classical to Next-Generation Strategies
Cid, R., & Bolívar, J. (2021) · Biomolecules · doi:10.3390/biom11081072 · Cited by 176
“Ionizable amine groups are positively charged at low pH, interacting with the anionic mRNA during particle formation and also facilitating membrane fusion during internalization [74,75].”
BackgroundMurphy, D. E., de Jong, O. G., Evers, M. J. W., Nurazizah, M., Schiffelers, R. M., & Vader, P. (2021) · Nano Letters · doi:10.1021/acs.nanolett.1c00094 · Cited by 169
Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads
Kulkarni, J. A., Witzigmann, D., Leung, J., van der Meel, R., Zaifman, J., Darjuan, M. M., Grisch-Chan, H. M., Thöny, B., Tam, Y. Y. C., & Cullis, P. R. (2019) · Nanoscale · doi:10.1039/c9nr02004g · Cited by 161
Ai, L., Li, Y., Zhou, L., Yao, W., Zhang, H., Hu, Z., Han, J., Wang, W., Wu, J., Xu, P., et al. (2023) · Cell Discovery · doi:10.1038/s41421-022-00517-9 · Cited by 148
Park, S., Choi, Y. K., Kim, S., Lee, J., & Im, W. (2021) · Journal of Chemical Information and Modeling · doi:10.1021/acs.jcim.1c00770 · Cited by 143
Throughput-scalable manufacturing of SARS-CoV-2 mRNA lipid nanoparticle vaccines
Shepherd, S. J., Han, X., Mukalel, A. J., El-Mayta, R. D., Thatte, A. S., Wu, J., Padilla, M. S., Alameh, M., Srikumar, N., Lee, D., et al. (2023) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2303567120 · Cited by 146
Nanomaterials for Therapeutic RNA Delivery
Han, X., Mitchell, M. J., & Nie, G. (2020) · Matter · doi:10.1016/j.matt.2020.09.020 · Cited by 139
Design of lipid-based nanoparticles for delivery of therapeutic nucleic acids
Mendonça, M. C. P., Kont, A., Kowalski, P. S., & O’Driscoll, C. M. (2023) · Drug Discovery Today · doi:10.1016/j.drudis.2023.103505 · Cited by 143
Mehraji, S., & DeVoe, D. L. (2024) · Lab on a Chip · doi:10.1039/d3lc00821e · Cited by 144
Current Status and Future Perspectives on MRNA Drug Manufacturing
Webb, C., Ip, S. Y., Bathula, N. V., Popova, P. G., Soriano, S. K. V., Ly, B.-H., Eryilmaz, B., Huu, V. A. N., Broadhead, R., Rabel, M., et al. (2022) · Molecular Pharmaceutics · doi:10.1021/acs.molpharmaceut.2c00010 · Cited by 134
Liu, K., Nilsson, R. G., Lázaro‐Ibáñez, E., Duàn, H., Miliotis, T., Strimfors, M., Lerche, M., Ribeiro, A. R. S., Ulander, J., Lindén, D., et al. (2023) · Nature Communications · doi:10.1038/s41467-023-39768-9 · Cited by 141
Microfluidic Manufacture of Lipid-Based Nanomedicines
Osouli-Bostanabad, K., Puliga, S., Serrano, D. R., Bucchi, A., Halbert, G., & Lalatsa, A. (2022) · Pharmaceutics · doi:10.3390/pharmaceutics14091940 · Cited by 137
Clinical delivery of circular RNA: Lessons learned from RNA drug development
Young, T. L., Wang, K. C., Varley, A., & Li, B. (2023) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2023.114826 · Cited by 130
Pattipeiluhu, R., Zeng, Y., Hendrix, M. M. R. M., Voets, I. K., Kros, A., & Sharp, T. H. (2024) · Nature Communications · doi:10.1038/s41467-024-45666-5 · Cited by 132
Optimal self-assembly of lipid nanoparticles (LNP) in a ring micromixer
Ripoll, M., Martin, E. A., Enot, M., Robbe, O., Rapisarda, C., Nicolai, M., Deliot, A., Tabeling, P., Authelin, J., Nakach, M., et al. (2022) · Scientific Reports · doi:10.1038/s41598-022-13112-5 · Cited by 105
mRNA delivery systems for cancer immunotherapy: Lipid nanoparticles and beyond
Sentí, M. E., del Valle, L. G., & Schiffelers, R. M. (2024) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2024.115190 · Cited by 118
Mechanisms and Barriers in Nanomedicine: Progress in the Field and Future Directions
Anchordoquy, T. J., Artzi, N., Balyasnikova, I. V., Barenholz, Y., La-Beck, N. M., Brenner, J. S., Chan, W. C. W., Decuzzi, P., Exner, A. A., Gabizón, A., et al. (2024) · ACS Nano · doi:10.1021/acsnano.4c00182 · Cited by 116
The Future of Tissue-Targeted Lipid Nanoparticle-Mediated Nucleic Acid Delivery
Kularatne, R. N., Crist, R. M., & Stern, S. T. (2022) · Pharmaceuticals · doi:10.3390/ph15070897 · Cited by 109
Xu, S., Xu, Y., Solek, N. C., Chen, J., Gong, F., Varley, A., Golubovic, A., Pan, A., Dong, S., Zheng, G., et al. (2024) · Advanced Materials · doi:10.1002/adma.202400307 · Cited by 112
Lipid Nanoparticle Technologies for Nucleic Acid Delivery: A Nanoarchitectonics Perspective
Ferhan, A. R., Park, S.-H., Park, H., Tae, H., Jackman, J. A., & Cho, N. (2022) · Advanced Functional Materials · doi:10.1002/adfm.202203669 · Cited by 103
Lipid nanoparticles: Composition, formulation, and application
Xu, S., Hu, Z., Song, F., Xu, Y. G., & Han, X. (2025) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2025.101463 · Cited by 112
“12 In this review, we first introduce the LNP components, highlighting their critical roles in encapsulation, stability, and delivery efficiency.”
Review of structural design guiding the development of lipid nanoparticles for nucleic acid delivery
Cárdenas, M., Campbell, R. A., Arteta, M. Y., Lawrence, M. J., & Sebastiani, F. (2023) · Current Opinion in Colloid & Interface Science · doi:10.1016/j.cocis.2023.101705 · Cited by 108
“…simple t-mixers, are used for scaling-up clinical and commercial manufacture of RNA LNP drug products since they are easy and cheap to scale and have the potential to form NPs with similar CQAs based on the organic NP precipitation principle depending on the geometry and the speed of mixing [66].”
MethodLipid Nanoparticles for Delivery of Therapeutic RNA Oligonucleotides
Moss, K. H., Popova, P. G., Hadrup, S. R., Astakhova, K., & Taskova, M. (2019) · Molecular Pharmaceutics · doi:10.1021/acs.molpharmaceut.8b01290 · Cited by 101
John, R., monpara, J., Swaminathan, S., & Kalhapure, R. S. (2024) · Pharmaceutics · doi:10.3390/pharmaceutics16010131 · Cited by 100
de Carvalho, B. G., Ceccato, B. T., Michelon, M., Han, S. W., & de la Torre, L. G. (2022) · Pharmaceutics · doi:10.3390/pharmaceutics14010141 · Cited by 97
“After reaching the cytoplasm, siRNA and miRNA interacts with the RNA-induced silencing complex (RISC), and then, the RISC is guided to the target mRNA, which is recognized and cleaved or blocked for translation [66,124].”
BackgroundMethodA Biopharmaceutical Perspective on Higher-Order Structure and Thermal Stability of mRNA Vaccines
Kloczewiak, M. A., Banks, J. M., Jin, L., & Brader, M. L. (2022) · Molecular Pharmaceutics · doi:10.1021/acs.molpharmaceut.2c00092 · Cited by 90
Advancements in mRNA Encoded Antibodies for Passive Immunotherapy
Deal, C. E., Carfı́, A., & Plante, O. J. (2021) · Vaccines · doi:10.3390/vaccines9020108 · Cited by 90
“trols to ensure reproducibility of manufacturing and stability [75].”
BackgroundMicrofluidic production of mRNA-loaded lipid nanoparticles for vaccine applications
Lopes, C., Cristóvão, J. S., Silvério, V., Lino, P. R., & Fonte, P. (2022) · Expert Opinion on Drug Delivery · doi:10.1080/17425247.2022.2135502 · Cited by 88
“small particles are only generated with Qratio > 30, leading to substantial dilution of the total solution [16].”
BackgroundBerger, M., Degey, M., Chain, J. L., Maquoi, E., Évrard, B., Lechanteur, A., & Piel, G. (2023) · Pharmaceutics · doi:10.3390/pharmaceutics15020597 · Cited by 92
“0 known to exhibit a neutral surface charge at physiological pH [18,28].”
BackgroundMethodPetersen, D. M. S., Chaudhary, N., Arral, M. L., Weiss, R. M., & Whitehead, K. A. (2023) · European Journal of Pharmaceutics and Biopharmaceutics · doi:10.1016/j.ejpb.2023.10.006 · Cited by 86
Opportunities and challenges in commercial pharmaceutical liposome applications
Jensen, G. M., & Hodgson, D. F. (2020) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2020.07.016 · Cited by 87
Doxorubicin-conjugated siRNA lipid nanoparticles for combination cancer therapy
Butowska, K., Han, X., Gong, N., El-Mayta, R. D., Haley, R. M., Xue, L., Zhong, W., Guo, W., Wang, K., & Mitchell, M. J. (2022) · Acta Pharmaceutica Sinica B · doi:10.1016/j.apsb.2022.07.011 · Cited by 84
Recent advances in the delivery and applications of nonviral CRISPR/Cas9 gene editing
Sinclair, F., Begum, A. A., Dai, C. C., Tóth, I., & Moyle, P. M. (2023) · Drug Delivery and Translational Research · doi:10.1007/s13346-023-01320-z · Cited by 91
“Current research is focused on improving the properties of LNPs with respect to cell-penetration, endosome escape, reducing toxicity, degradation, and improving long-term storage stability [47, 48].”
BackgroundZukancic, D., Suys, E. J. A., Pilkington, E. H., Algarni, A., Al-Wassiti, H. A., & Truong, N. P. (2020) · Pharmaceutics · doi:10.3390/pharmaceutics12111068 · Cited by 81
“In this work, LNPs encapsulating pDNA were prepared using a scalable, rapid, and reproducible microfluidic mixing technology (NanoAssemblr®) [40].”
MethodO’Brien, M. N., Costa, A. P., Cebrero, Y. M., Joseph, J., Sarode, A., Zang, N., Kim, L. J., Hofmann, K., Wang, S., Goyon, A., et al. (2023) · Molecular Pharmaceutics · doi:10.1021/acs.molpharmaceut.3c00390 · Cited by 78
Cytotoxic effects of nanoparticles assessed in vitro and in vivo.
Cha, K. E., & Myung, H. (2007) · PubMed · Cited by 75
The mRNA Vaccine Revolution: COVID-19 Has Launched the Future of Vaccinology
Ye, Z., Harmon, J., Ni, W., Li, Y., Wich, D., & Xu, Q. (2023) · ACS Nano · doi:10.1021/acsnano.2c12584 · Cited by 72
Spontaneous, solvent-free entrapment of siRNA within lipid nanoparticles
Kulkarni, J. A., Thomson, S. B., Zaifman, J., Leung, J., Wagner, P. K., Hill, A. M., Tam, Y. Y. C., Cullis, P. R., Petkau, T. L., & Leavitt, B. R. (2020) · Nanoscale · doi:10.1039/d0nr06816k · Cited by 73
Leveraging Biological Buffers for Efficient Messenger RNA Delivery via Lipid Nanoparticles
Henderson, M. I., Eygeris, Y., Jozić, A., Herrera‐Barrera, M., & Sahay, G. (2022) · Molecular Pharmaceutics · doi:10.1021/acs.molpharmaceut.2c00587 · Cited by 71
Development of a high-throughput platform for screening lipid nanoparticles for mRNA delivery
Cui, L., Pereira, S. G. T., Sonzini, S., van Pelt, S., Romanelli, S. M., Liang, L., Ulkoski, D., Krishnamurthy, V. R., Brannigan, E., Brankin, C., et al. (2022) · Nanoscale · doi:10.1039/d1nr06858j · Cited by 72
Lipid nanoparticle-mediated messenger RNA delivery for ex vivo engineering of natural killer cells
Douka, S., Brandenburg, L. E., Casadidio, C., Walther, J., Garcia, B. B. M., Spanholtz, J., Raimo, M., Hennink, W. E., Mastrobattista, E., & Caiazzo, M. (2023) · Journal of Controlled Release · doi:10.1016/j.jconrel.2023.08.014 · Cited by 70
Cai, X., Drummond, C. J., Zhai, J., & Tran, N. L. (2024) · Advanced Functional Materials · doi:10.1002/adfm.202404234 · Cited by 75
Wang, Z., Cui, K., Costabel, U., & Zhang, X. (2022) · Exploration · doi:10.1002/exp.20210082 · Cited by 68
Structure and Morphology of Lipid Nanoparticles for Nucleic Acid Drug Delivery: A Review
Udepurkar, A. P., Devos, C., Sagmeister, P., Destro, F., Inguva, P., Ahmadi, S., Boulais, É., Quan, Y., Braatz, R. D., & Myerson, A. S. (2025) · ACS Nano · doi:10.1021/acsnano.4c18274 · Cited by 74
Fan, Y., Yen, C., Lin, H.-C., Hou, W., Estevez, A., Sarode, A., Goyon, A., Bian, J., Lin, J., Koenig, S. G., et al. (2021) · International Journal of Pharmaceutics · doi:10.1016/j.ijpharm.2021.120392 · Cited by 68
Fluoroalkane modified cationic polymers for personalized mRNA cancer vaccines
Li, J., Wu, Y., Xiang, J., Wang, H., Zhuang, Q., Wei, T., Cao, Z., Gu, Q., Liu, Z., & Peng, R. (2022) · Chemical Engineering Journal · doi:10.1016/j.cej.2022.140930 · Cited by 64
Composition of lipid nanoparticles for targeted delivery: application to mRNA therapeutics
Vasileva, O., Zaborova, O. V., Shmykov, B., Ivanov, R. A., & Reshetnikov, V. V. (2024) · Frontiers in Pharmacology · doi:10.3389/fphar.2024.1466337 · Cited by 72
Advanced manufacturing of nanoparticle formulations of drugs and biologics using microfluidics
Shen, Y., Gwak, H., & Han, B. (2023) · The Analyst · doi:10.1039/d3an01739g · Cited by 65
Intracellular Routing and Recognition of Lipid-Based mRNA Nanoparticles
Delehedde, C., Even, L., Midoux, P., Pichon, C., & Perche, F. (2021) · Pharmaceutics · doi:10.3390/pharmaceutics13070945 · Cited by 60
“Decreasing mRNA leakage from LNPs and improving their circulation time have been the focus of extensive research (reviewed in [25]).”
BackgroundInfluentialCharacterization of lipid-based nanomedicines at the single-particle level
Chen, C., Chen, C., Li, Y., Gu, R., & Yan, X. (2022) · Fundamental Research · doi:10.1016/j.fmre.2022.09.011 · Cited by 55
“…in revealing heterogeneity and ensuring quality control of the LBNMs. Cryo-TEM is also applicable to confirm the electron dense core of LNPs, as well as to investigate the impact of cationic lipids, pH value and drug-to-lipid ratio on the size distribution and morphology of LNPs [ 54 , 55 ].”
BackgroundPittiu, A., Pannuzzo, M., Casula, L., Pireddu, R., Valenti, D., Cardia, M. C., Lai, F., Rosa, A., Sinico, C., & Schlich, M. (2024) · International Journal of Pharmaceutics · doi:10.1016/j.ijpharm.2024.124641 · Cited by 52
“The staggered herringbone microchip has two inlet channels, one for the organic phase containing lipids and one for the aqueous buffer, which converge in one central channel where the mixing happens, leading to the self-assembly of lipids into vesicular structures (Evers et al., 2018).”
Design Strategies for and Stability of mRNA–Lipid Nanoparticle COVID-19 Vaccines
Liu, T., Tian, Y., Zheng, A., & Cui, C. (2022) · Polymers · doi:10.3390/polym14194195 · Cited by 54
“With the improvement of technology, large-scale production, which proves difficult for obtaining a greater R e tion efficiency and uniform particle siz , s generally no l nger practiced [92,93].”
BackgroundMethodHigh‐Precision Synthesis of RNA‐Loaded Lipid Nanoparticles for Biomedical Applications
Seo, H., Jeon, L., Kwon, J., & Lee, H. (2023) · Advanced Healthcare Materials · doi:10.1002/adhm.202203033 · Cited by 54
Polysarcosine-Functionalized mRNA Lipid Nanoparticles Tailored for Immunotherapy
Wilhelmy, C., Keil, I. S., Uebbing, L., Schroer, M. A., Franke, D., Nawroth, T., Barz, M., Şahin, U., Haas, H., Diken, M., et al. (2023) · Pharmaceutics · doi:10.3390/pharmaceutics15082068 · Cited by 51
“Currently approved LNP formulations are characterized by a very specific lipid composition, consisting of four primary components: ionizable-cationic lipid, cholesterol, helper lipid and a PEG stealth moiety [10,57].”
BackgroundLipid nanoparticles for RNA delivery: Self-assembling vs driven-assembling strategies
Nele, V., Campani, V., Moosavian, S. A., & Rosa, G. D. (2024) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2024.115291 · Cited by 58
Nel, A. E., & Miller, J. F. (2021) · ACS Nano · doi:10.1021/acsnano.1c01845 · Cited by 50
Microfluidic Formulation of DNA-Loaded Multicomponent Lipid Nanoparticles for Gene Delivery
Quagliarini, E., Renzi, S., Digiacomo, L., Giulimondi, F., Sartori, B., Amenitsch, H. W., Tassinari, V., Masuelli, L., Bei, R., Cui, L., et al. (2021) · Pharmaceutics · doi:10.3390/pharmaceutics13081292 · Cited by 51
“Thus, new production strategies to encapsulate NAs need to deal with the issues of reproducibility, scalability of production and encapsulation efficiency [12].”
BackgroundThe journey of a lifetime — development of Pfizer’s COVID-19 vaccine
Thorn, C. R., Sharma, D., Combs, R. G., Bhujbal, S. V., Romine, J., Zheng, X., Sunasara, K., & Badkar, A. V. (2022) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2022.102803 · Cited by 49
Microfluidic technologies for lipid vesicle generation
Yu, C., Hay, C. D., Mahuttanatan, S. M., Hindley, J. W., Ces, O., & Elani, Y. (2024) · Lab on a Chip · doi:10.1039/d4lc00380b · Cited by 53
Microfluidic synthesis of nanomaterials for biomedical applications
Huang, Y., Liu, C., Feng, Q., & Sun, J. (2023) · Nanoscale Horizons · doi:10.1039/d3nh00217a · Cited by 52
CRISPR-Cas9 Gene Therapy: Non-Viral Delivery and Stimuli-Responsive Nanoformulations
Lee, H., Rho, W., Kim, Y.-H., Chang, H., & Jun, B. (2025) · Molecules · doi:10.3390/molecules30030542 · Cited by 59
“Structurally, LNPs are spherical vesicles composed of lipids with hydrophilic head groups and hydrophobic tail groups, which self-assemble into nanostructures in aqueous environments [70].”
Learning from the future: towards continuous manufacturing of nanomaterials
VandenBerg, M. A., Dong, X., Smith, W. C., Tian, G., Stephens, O., O’Connor, T. F., & Xu, X. (2025) · AAPS Open · doi:10.1186/s41120-025-00111-9 · Cited by 49
“…that have been adopted in the last few decades as improvements over such methods: chaotic advection to enhance mixing through microchannel geometry, and hydrodynamic flow focusing which adds convective mixing contributions (Evers MJW et al 2018; Hamdallah SIZ et al 2020), depicted in Fig.”
mRNA Delivery: Challenges and Advances through Polymeric Soft Nanoparticles
Adlsadabad, S. Y., Hanrahan, J. W., & Kakkar, A. K. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25031739 · Cited by 47
“[55]”
Lipid nanoparticles as the drug carrier for targeted therapy of hepatic disorders
Chu, R., Wang, Y., Kong, J., Pan, T. S., Yang, Y., & He, J. F. (2024) · Journal of Materials Chemistry B · doi:10.1039/d3tb02766j · Cited by 49
Nanoparticles as Drug Delivery Systems of RNAi in Cancer Therapy
Li, D., Gao, C., Kuang, M., Xu, M., Wang, B., Luo, Y., Teng, L., & Xie, J. (2021) · Molecules · doi:10.3390/molecules26082380 · Cited by 45
Dhumal, D. M., Lan, W., Ding, L., Jiang, Y., Lyu, Z., Laurini, E., Marson, D., Tintaru, A., Dusetti, N., Giorgio, S., et al. (2020) · Nano Research · doi:10.1007/s12274-020-3216-8 · Cited by 46
Recent Advances in Drug Delivery System Fabricated by Microfluidics for Disease Therapy
Jia, F., Gao, Y., & Wang, H. (2022) · Bioengineering · doi:10.3390/bioengineering9110625 · Cited by 40
“In this case, the rapidly changing polarity supersaturates lipids, causing the lipids to self-assemble to form liposomes or LNPs (Figure 5a) [87].”
BackgroundNon-fouling flow reactors for nanomaterial synthesis
Besenhard, M. O., Pal, S., Gkogkos, G., & Gavriilidis, A. (2023) · Reaction Chemistry & Engineering · doi:10.1039/d2re00412g · Cited by 41
Lu, J., Atochina‐Vasserman, E. N., Maurya, D. S., Shalihin, M. I., Zhang, D., Chenna, S. S., Adamson, J., Liu, M., Shah, H. U. R., Shah, H., et al. (2022) · Pharmaceutics · doi:10.3390/pharmaceutics15061572 · Cited by 40
Seder, I., Zheng, T., Zhang, J., Rojas, C. C., Helalat, S. H., Coronel‐Tellez, R. H., & Sun, Y. (2024) · Nano Letters · doi:10.1021/acs.nanolett.3c05057 · Cited by 40
Are inhaled mRNA vaccines safe and effective? A review of preclinical studies
Jansen, E. M., Frijlink, H. W., Hinrichs, W. L. J., & Ruigrok, M. J. R. (2022) · Expert Opinion on Drug Delivery · doi:10.1080/17425247.2022.2131767 · Cited by 37
“These findings are remarkable, considering the fact that the uptake of naked mRNA was shown to be negligible in vitro, owing to its large size and negative charge as well as susceptibility to nucleases [54,64].”
BackgroundContinuous Production of the Nanoscale Liposome in a Double Flow-Focusing Microfluidic Device
Aghaei, H., & Nazar, A. R. S. (2019) · Industrial & Engineering Chemistry Research · doi:10.1021/acs.iecr.9b04079 · Cited by 42
Metadata
- Authors
- Evers, M. J. W., Kulkarni, J. A., van der Meel, R., Cullis, P. R., Vader, P., & Schiffelers, R. M.
- Affiliations
- University Medical Center Utrecht (NL)
- University of British Columbia (CA)
- Department of Clinical Chemistry and Haematology University Medical Center Utrecht Heidelberglaan 100 3584 CX Utrecht The Netherlands
- Corresponding author
- Schiffelers, Raymond M.
- Venue
- Small Methods
- Publisher
- Wiley
- Published
- 2018-04-26
- Volume
- 2
- Issue
- 9
- ISSN
- 2366-9608 (ISSN-L)
- OpenAlex
- W2799813152
- Cited by
- 332
- References
- 153
- Citations by year
- 2026: 52 · 2025: 55 · 2024: 56 · 2023: 58 · 2022: 52 · 2021: 32 · 2020: 15 · 2019: 9 · 2018: 2
- FWCI
- 5.15
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- public-domain
- onlinelibrary.wiley.com/termsAndConditions#vor (vor)
- Locations
- Small Methods (publishedVersion, public-domain)
- TU/e Research Portal (submittedVersion)
- Utrecht University Repository (Utrecht University) (submittedVersion, other-oa)
- Zenodo (CERN European Organization for Nuclear Research) (submittedVersion, other-oa)
- Utrecht University Repository (Utrecht University) (submittedVersion)
- Funders
- H2020 LEIT Nanotechnologies (721058, # 14385, # 13667)
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek
- Canadian Institutes of Health Research (148469)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Advancements in Transdermal Drug Delivery
- Keywords
- lipid nanoparticles, siRNA delivery, non-viral gene delivery, pharmacokinetics, gene silencing, mRNA delivery, CRISPR/Cas9 delivery, scalable fabrication, experimental design, nucleic acid carriers
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
