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The most striking example of the leap in the development of mRNA technologies was the mass vaccination against COVID-19 during the pandemic. The emergence of large-scale technology and positive experience of mRNA immunization sparked the development of antiviral and anti-cancer mRNA vaccines as well as therapeutic mRNA agents for genetic and other diseases. To facilitate mRNA delivery, lipid nanoparticles (LNPs) have been successfully employed. However, the diverse use of mRNA therapeutic approaches requires the development of adaptable LNP delivery systems that can control the kinetics of mRNA uptake and expression in target cells. Here, we report effective mRNA delivery into cultured mammalian cells (HEK293T, HeLa, DC2.4) and living mouse muscle tissues by liposomes containing either 1,26-bis(cholest-5-en-3\u03b2-yloxycarbonylamino)-7,11,16,20-tetraazahexacosane tetrahydrochloride (2X3) or the newly applied 1,30-bis(cholest-5-en-3\u03b2-yloxycarbonylamino)-9,13,18,22-tetraaza-3,6,25,28-tetraoxatriacontane tetrahydrochloride (2X7) cationic lipids. Using end-point and real-time monitoring of Fluc mRNA expression, we showed that these LNPs exhibited an unusually delayed (of over 10 h in the case of the 2X7-based system) but had highly efficient and prolonged reporter activity in cells. Accordingly, both LNP formulations decorated with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000) provided efficient luciferase production in mice, peaking on day 3 after intramuscular injection. Notably, the bioluminescence was observed only at the site of injection in caudal thigh muscles, thereby demonstrating local expression of the model gene of interest. The developed mRNA delivery systems hold promise for prophylactic applications, where sustained synthesis of defensive proteins is required, and open doors to new possibilities in mRNA-based therapies.<\/jats:p>","DOI":"10.3390\/pharmaceutics16050684","type":"journal-article","created":{"date-parts":[[2024,5,20]],"date-time":"2024-05-20T11:06:41Z","timestamp":1716203201000},"page":"684","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Novel Efficient Lipid-Based Delivery Systems Enable a Delayed Uptake and Sustained Expression of mRNA in Human Cells and Mouse Tissues"],"prefix":"10.3390","volume":"16","author":[{"given":"Artem G.","family":"Fedorovskiy","sequence":"first","affiliation":[{"name":"Belozersky Institute of Physico-Chemical Biology, Department of Materials Science, Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119234 Moscow, Russia"},{"name":"Lomonosov Institute of Fine Chemical Technologies, MIREA-Russian Technological University, 119571 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Denis N.","family":"Antropov","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2704-3604","authenticated-orcid":false,"given":"Anton S.","family":"Dome","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6591-950X","authenticated-orcid":false,"given":"Pavel A.","family":"Puchkov","sequence":"additional","affiliation":[{"name":"Lomonosov Institute of Fine Chemical Technologies, MIREA-Russian Technological University, 119571 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daria M.","family":"Makarova","sequence":"additional","affiliation":[{"name":"Lomonosov Institute of Fine Chemical Technologies, MIREA-Russian Technological University, 119571 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9724-695X","authenticated-orcid":false,"given":"Maria V.","family":"Konopleva","sequence":"additional","affiliation":[{"name":"Belozersky Institute of Physico-Chemical Biology, Department of Materials Science, Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119234 Moscow, Russia"},{"name":"Lomonosov Institute of Fine Chemical Technologies, MIREA-Russian Technological University, 119571 Moscow, Russia"},{"name":"Federal State Budget Institution \u201cNational Research Centre for Epidemiology and Microbiology Named after Honorary Academician N.F. Gamaleya\u201d of the Ministry of Health of the Russian Federation, 123098 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anastasiya M.","family":"Matveeva","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eugenia A.","family":"Panova","sequence":"additional","affiliation":[{"name":"Belozersky Institute of Physico-Chemical Biology, Department of Materials Science, Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119234 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3727-4905","authenticated-orcid":false,"given":"Elena V.","family":"Shmendel","sequence":"additional","affiliation":[{"name":"Lomonosov Institute of Fine Chemical Technologies, MIREA-Russian Technological University, 119571 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5372-1325","authenticated-orcid":false,"given":"Mikhail A.","family":"Maslov","sequence":"additional","affiliation":[{"name":"Lomonosov Institute of Fine Chemical Technologies, MIREA-Russian Technological University, 119571 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1774-8475","authenticated-orcid":false,"given":"Sergey E.","family":"Dmitriev","sequence":"additional","affiliation":[{"name":"Belozersky Institute of Physico-Chemical Biology, Department of Materials Science, Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119234 Moscow, Russia"},{"name":"Federal State Budget Institution \u201cNational Research Centre for Epidemiology and Microbiology Named after Honorary Academician N.F. Gamaleya\u201d of the Ministry of Health of the Russian Federation, 123098 Moscow, Russia"},{"name":"Moscow Institute of Physics and Technology, 141701 Dolgoprudny, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3393-3192","authenticated-orcid":false,"given":"Grigory A.","family":"Stepanov","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8381-1890","authenticated-orcid":false,"given":"Oleg V.","family":"Markov","sequence":"additional","affiliation":[{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1038\/s41392-022-01007-w","article-title":"mRNA-based therapeutics: Powerful and versatile tools to combat diseases","volume":"7","author":"Qin","year":"2022","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/s12943-021-01335-5","article-title":"mRNA vaccine for cancer immunotherapy","volume":"20","author":"Miao","year":"2021","journal-title":"Mol. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"840","DOI":"10.1038\/s41587-022-01294-2","article-title":"The clinical progress of mRNA vaccines and immunotherapies","volume":"40","author":"Barbier","year":"2022","journal-title":"Nat. Biotechnol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1586\/14760584.2015.973859","article-title":"mRNA transcript therapy","volume":"14","author":"Weissman","year":"2015","journal-title":"Expert Rev. Vaccines"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1038\/nrd4278","article-title":"mRNA-based therapeutics--developing a new class of drugs","volume":"13","author":"Sahin","year":"2014","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Hauck, E.S., and Hecker, J.G. (2022). Non-Viral Delivery of RNA Gene Therapy to the Central Nervous System. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14010165"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1056\/NEJMoa2035389","article-title":"Efficacy and Safety of the mRNA-1273 SARS-CoV-2 Vaccine","volume":"384","author":"Baden","year":"2021","journal-title":"N. Engl. J. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1038\/s41586-020-2622-0","article-title":"SARS-CoV-2 mRNA vaccine design enabled by prototype pathogen preparedness","volume":"586","author":"Corbett","year":"2020","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2603","DOI":"10.1056\/NEJMoa2034577","article-title":"Safety and Efficacy of the BNT162b2 mRNA COVID-19 Vaccine","volume":"383","author":"Polack","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1038\/s41573-021-00283-5","article-title":"mRNA vaccines for infectious diseases: Principles, delivery and clinical translation","volume":"20","author":"Chaudhary","year":"2021","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1434","DOI":"10.15252\/emmm.201707678","article-title":"mRNA mediates passive vaccination against infectious agents, toxins, and tumors","volume":"9","author":"Thran","year":"2017","journal-title":"EMBO Mol. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1186\/s12967-019-1804-8","article-title":"How mRNA therapeutics are entering the monoclonal antibody field","volume":"17","author":"Roose","year":"2019","journal-title":"J. Transl. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1038\/nrd.2017.243","article-title":"mRNA vaccines\u2014A new era in vaccinology","volume":"17","author":"Pardi","year":"2018","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"16982","DOI":"10.1021\/acsnano.1c04996","article-title":"Lipid Nanoparticles\u2014From Liposomes to mRNA Vaccine Delivery, a Landscape of Research Diversity and Advancement","volume":"15","author":"Tenchov","year":"2021","journal-title":"ACS Nano"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1078","DOI":"10.1038\/s41578-021-00358-0","article-title":"Lipid nanoparticles for mRNA delivery","volume":"6","author":"Hou","year":"2021","journal-title":"Nat. Rev. Mater."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1038\/s41576-021-00439-4","article-title":"Drug delivery systems for RNA therapeutics","volume":"23","author":"Paunovska","year":"2022","journal-title":"Nat. Rev. Genet."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Huang, P., Deng, H., Wang, C., Zhou, Y., and Chen, X. (2023). Cellular Trafficking of Nanotechnology-Mediated mRNA Delivery. Adv. Mater., e2307822.","DOI":"10.1002\/adma.202307822"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.addr.2019.08.004","article-title":"Brief update on endocytosis of nanomedicines","volume":"144","author":"Patel","year":"2019","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"114325","DOI":"10.1016\/j.addr.2022.114325","article-title":"In vivo fate and intracellular trafficking of vaccine delivery systems","volume":"186","author":"Lee","year":"2022","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Szewczyk-Roszczenko, O.K., Roszczenko, P., Shmakova, A., Finiuk, N., Holota, S., Lesyk, R., Bielawska, A., Vassetzky, Y., and Bielawski, K. (2023). The Chemical Inhibitors of Endocytosis: From Mechanisms to Potential Clinical Applications. Cells, 12.","DOI":"10.3390\/cells12182312"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Delehedde, C., Even, L., Midoux, P., Pichon, C., and Perche, F. (2021). Intracellular Routing and Recognition of Lipid-Based mRNA Nanoparticles. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13070945"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1038\/nbt.2634","article-title":"A window onto siRNA delivery","volume":"31","author":"Wang","year":"2013","journal-title":"Nat. Biotechnol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1038\/nbt.3298","article-title":"Visualizing lipid-formulated siRNA release from endosomes and target gene knockdown","volume":"33","author":"Wittrup","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"638","DOI":"10.1038\/nbt.2612","article-title":"Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape","volume":"31","author":"Gilleron","year":"2013","journal-title":"Nat. Biotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Paramasivam, P., Franke, C., Stoter, M., Hoijer, A., Bartesaghi, S., Sabirsh, A., Lindfors, L., Arteta, M.Y., Dahlen, A., and Bak, A. (2022). Endosomal escape of delivered mRNA from endosomal recycling tubules visualized at the nanoscale. J. Cell Biol., 221.","DOI":"10.1083\/jcb.202110137"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1038\/nbt.2614","article-title":"Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling","volume":"31","author":"Sahay","year":"2013","journal-title":"Nat. Biotechnol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1583","DOI":"10.1093\/nar\/gkad023","article-title":"Simultaneous inhibition of endocytic recycling and lysosomal fusion sensitizes cells and tissues to oligonucleotide therapeutics","volume":"51","author":"Finicle","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4333","DOI":"10.1038\/s41467-019-12275-6","article-title":"Linkage between endosomal escape of LNP-mRNA and loading into EVs for transport to other cells","volume":"10","author":"Maugeri","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2100","DOI":"10.1038\/mt.2016.179","article-title":"The Niemann-Pick C1 Inhibitor NP3.47 Enhances Gene Silencing Potency of Lipid Nanoparticles Containing siRNA","volume":"24","author":"Wang","year":"2016","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3654","DOI":"10.1021\/acs.molpharmaceut.0c00618","article-title":"Nanomedicine-Based Approaches for mRNA Delivery","volume":"17","author":"Uchida","year":"2020","journal-title":"Mol. Pharm."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1509","DOI":"10.1016\/j.ymthe.2018.03.010","article-title":"A Novel Amino Lipid Series for mRNA Delivery: Improved Endosomal Escape and Sustained Pharmacology and Safety in Non-human Primates","volume":"26","author":"Sabnis","year":"2018","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"114416","DOI":"10.1016\/j.addr.2022.114416","article-title":"The role of lipid components in lipid nanoparticles for vaccines and gene therapy","volume":"188","author":"Kulkarni","year":"2022","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1021\/acs.accounts.1c00544","article-title":"Chemistry of Lipid Nanoparticles for RNA Delivery","volume":"55","author":"Eygeris","year":"2022","journal-title":"Acc. Chem. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.addr.2016.01.022","article-title":"The role of helper lipids in lipid nanoparticles (LNPs) designed for oligonucleotide delivery","volume":"99","author":"Cheng","year":"2016","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"12181","DOI":"10.1021\/acs.chemrev.1c00244","article-title":"Lipids and Lipid Derivatives for RNA Delivery","volume":"121","author":"Zhang","year":"2021","journal-title":"Chem. Rev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"5711","DOI":"10.1021\/acs.nanolett.7b02664","article-title":"Boosting Intracellular Delivery of Lipid Nanoparticle-Encapsulated mRNA","volume":"17","author":"Patel","year":"2017","journal-title":"Nano Lett."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1038\/nbt.1602","article-title":"Rational design of cationic lipids for siRNA delivery","volume":"28","author":"Semple","year":"2010","journal-title":"Nat. Biotechnol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"7300","DOI":"10.1021\/acs.nanolett.5b02497","article-title":"Optimization of Lipid Nanoparticle Formulations for mRNA Delivery in Vivo with Fractional Factorial and Definitive Screening Designs","volume":"15","author":"Kauffman","year":"2015","journal-title":"Nano Lett."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1038\/s41563-020-00886-0","article-title":"Membrane-destabilizing ionizable phospholipids for organ-selective mRNA delivery and CRISPR-Cas gene editing","volume":"20","author":"Liu","year":"2021","journal-title":"Nat. Mater."},{"key":"ref_40","first-page":"E3351","article-title":"Successful reprogramming of cellular protein production through mRNA delivered by functionalized lipid nanoparticles","volume":"115","author":"Kjellman","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e2301067120","DOI":"10.1073\/pnas.2301067120","article-title":"Lipid nanoparticle topology regulates endosomal escape and delivery of RNA to the cytoplasm","volume":"120","author":"Zheng","year":"2023","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Gomez-Aguado, I., Rodriguez-Castejon, J., Vicente-Pascual, M., Rodriguez-Gascon, A., Solinis, M.A., and Del Pozo-Rodriguez, A. (2020). Nanomedicines to Deliver mRNA: State of the Art and Future Perspectives. Nanomaterials, 10.","DOI":"10.3390\/nano10020364"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1038\/s41467-020-14527-2","article-title":"Naturally-occurring cholesterol analogues in lipid nanoparticles induce polymorphic shape and enhance intracellular delivery of mRNA","volume":"11","author":"Patel","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"4375","DOI":"10.1093\/nar\/gkz214","article-title":"Differential uptake, kinetics and mechanisms of intracellular trafficking of next-generation antisense oligonucleotides across human cancer cell lines","volume":"47","author":"Linnane","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_45","first-page":"362","article-title":"Correlation of mRNA delivery timing and protein expression in lipid-based transfection","volume":"11","author":"Reiser","year":"2019","journal-title":"Integr. Biol. Quant. Biosci. Nano Macro."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"5309","DOI":"10.1093\/nar\/gkx231","article-title":"Intra-endosomal trafficking mediated by lysobisphosphatidic acid contributes to intracellular release of phosphorothioate-modified antisense oligonucleotides","volume":"45","author":"Wang","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.ijpharm.2010.01.019","article-title":"Lipid-mediated delivery of RNA is more efficient than delivery of DNA in non-dividing cells","volume":"389","author":"Zou","year":"2010","journal-title":"Int. J. Pharm."},{"key":"ref_48","first-page":"7314","article-title":"Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides","volume":"44","author":"Wang","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1038\/nature15377","article-title":"Dynamic m(6)A mRNA methylation directs translational control of heat shock response","volume":"526","author":"Zhou","year":"2015","journal-title":"Nature"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1016\/j.nano.2013.11.008","article-title":"Single-cell mRNA transfection studies: Delivery, kinetics and statistics by numbers","volume":"10","author":"Leonhardt","year":"2014","journal-title":"Nanomed. Nanotechnol. Biol. Med."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"3767","DOI":"10.1021\/nn3049494","article-title":"Mechanism of polyplex- and lipoplex-mediated delivery of nucleic acids: Real-time visualization of transient membrane destabilization without endosomal lysis","volume":"7","author":"Hoekstra","year":"2013","journal-title":"ACS Nano"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.jconrel.2011.11.023","article-title":"Novel cholesterol spermine conjugates provide efficient cellular delivery of plasmid DNA and small interfering RNA","volume":"160","author":"Maslov","year":"2012","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.jconrel.2011.11.034","article-title":"Novel cationic liposomes provide highly efficient delivery of DNA and RNA into dendritic cell progenitors and their immature offsets","volume":"160","author":"Markov","year":"2012","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Vysochinskaya, V., Shishlyannikov, S., Zabrodskaya, Y., Shmendel, E., Klotchenko, S., Dobrovolskaya, O., Gavrilova, N., Makarova, D., Plotnikova, M., and Elpaeva, E. (2022). Influence of Lipid Composition of Cationic Liposomes 2X3-DOPE on mRNA Delivery into Eukaryotic Cells. Pharmaceutics, 15.","DOI":"10.3390\/pharmaceutics15010008"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"3284","DOI":"10.1016\/j.bmcl.2017.06.026","article-title":"Spacer structure and hydrophobicity influences transfection activity of novel polycationic gemini amphiphiles","volume":"27","author":"Puchkov","year":"2017","journal-title":"Bioorganic Med. Chem. Lett."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"7107","DOI":"10.1038\/srep07107","article-title":"The role of the helper lipid on the DNA transfection efficiency of lipopolyplex formulations","volume":"4","author":"Du","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"253","DOI":"10.3892\/mmr.2022.12769","article-title":"Optimal combination of cationic lipid and phospholipid in cationic liposomes for gene knockdown in breast cancer cells and mouse lung using siRNA lipoplexes","volume":"26","author":"Hattori","year":"2022","journal-title":"Mol. Med. Rep."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Ewert, K.K., Scodeller, P., Simon-Gracia, L., Steffes, V.M., Wonder, E.A., Teesalu, T., and Safinya, C.R. (2021). Cationic Liposomes as Vectors for Nucleic Acid and Hydrophobic Drug Therapeutics. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13091365"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1038\/s41589-020-0578-x","article-title":"Tetracenomycin X inhibits translation by binding within the ribosomal exit tunnel","volume":"16","author":"Osterman","year":"2020","journal-title":"Nat. Chem. Biol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"15473","DOI":"10.1038\/s41598-020-72399-4","article-title":"Translation elongation factor 2 depletion by siRNA in mouse liver leads to mTOR-independent translational upregulation of ribosomal protein genes","volume":"10","author":"Gerashchenko","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1098302","DOI":"10.3389\/fimmu.2023.1098302","article-title":"Single-domain antibody delivery using an mRNA platform protects against lethal doses of botulinum neurotoxin A","volume":"14","author":"Panova","year":"2023","journal-title":"Front. Immunol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"724","DOI":"10.1134\/S1068162019010084","article-title":"Optimization of the Technology for the Preparation of Cationic Liposomes for the Delivery of Nucleic Acids","volume":"44","author":"Luneva","year":"2018","journal-title":"Russ. J. Bioorganic Chem."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1007\/s11172-010-0071-x","article-title":"Synthesis of polycationic lipids based on cholesterol and spermine","volume":"59","author":"Petukhov","year":"2010","journal-title":"Russ. Chem. Bull."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1038\/s41587-019-0164-5","article-title":"Human 5\u2032 UTR design and variant effect prediction from a massively parallel translation assay","volume":"37","author":"Sample","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1016\/j.ymthe.2018.12.011","article-title":"Improving mRNA-Based Therapeutic Gene Delivery by Expression-Augmenting 3\u2032 UTRs Identified by Cellular Library Screening","volume":"27","author":"Poleganov","year":"2019","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"37905","DOI":"10.1038\/srep37905","article-title":"Four translation initiation pathways employed by the leaderless mRNA in eukaryotes","volume":"6","author":"Akulich","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"1786","DOI":"10.1134\/S000629792311010X","article-title":"A Solitary Stalled 80S Ribosome Prevents mRNA Recruitment to Stress Granules","volume":"88","author":"Fedorovskiy","year":"2023","journal-title":"Biochemistry. Biokhimiia"},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Markov, O.V., Sen\u2019kova, A.V., Mohamed, I.S., Shmendel, E.V., Maslov, M.A., Oshchepkova, A.L., Brenner, E.V., Mironova, N.L., and Zenkova, M.A. (2022). Dendritic Cell-Derived Artificial Microvesicles Inhibit RLS(40) Lymphosarcoma Growth in Mice via Stimulation of Th1\/Th17 Immune Response. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14112542"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1016\/0005-2736(95)80016-9","article-title":"The role of dioleoyl phosphatidylethanolamine in cationic liposome mediated gene transfer","volume":"1235","author":"Farhood","year":"1995","journal-title":"Biochim. Et Biophys. Acta"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"21733","DOI":"10.1039\/C9NR09347H","article-title":"On the role of helper lipids in lipid nanoparticle formulations of siRNA","volume":"11","author":"Kulkarni","year":"2019","journal-title":"Nanoscale"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1126\/science.281.5373.78","article-title":"An inverted hexagonal phase of cationic liposome-DNA complexes related to DNA release and delivery","volume":"281","author":"Koltover","year":"1998","journal-title":"Science"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1016\/j.omtm.2020.07.015","article-title":"Barriers and Strategies of Cationic Liposomes for Cancer Gene Therapy","volume":"18","author":"Liu","year":"2020","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1039\/C8MD00515J","article-title":"Diversity of PEGylation methods of liposomes and their influence on RNA delivery","volume":"10","author":"Nosova","year":"2019","journal-title":"MedChemComm"},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Berger, M., Degey, M., Leblond Chain, J., Maquoi, E., Evrard, B., Lechanteur, A., and Piel, G. (2023). Effect of PEG Anchor and Serum on Lipid Nanoparticles: Development of a Nanoparticles Tracking Method. Pharmaceutics, 15.","DOI":"10.3390\/pharmaceutics15020597"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/j.jconrel.2021.05.021","article-title":"Impact of lipid nanoparticle size on mRNA vaccine immunogenicity","volume":"335","author":"Hassett","year":"2021","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Gaspar, R., Coelho, F., and Silva, B.F.B. (2020). Lipid-Nucleic Acid Complexes: Physicochemical Aspects and Prospects for Cancer Treatment. Molecules, 25.","DOI":"10.3390\/molecules25215006"},{"key":"ref_77","doi-asserted-by":"crossref","unstructured":"Carrasco, M.J., Alishetty, S., Alameh, M.G., Said, H., Wright, L., Paige, M., Soliman, O., Weissman, D., Cleveland, T.E., and Grishaev, A. (2021). Ionization and structural properties of mRNA lipid nanoparticles influence expression in intramuscular and intravascular administration. Commun. Biol., 4.","DOI":"10.1038\/s42003-021-02441-2"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"8698","DOI":"10.1021\/acs.jpcb.5b02891","article-title":"Microfluidic Mixing: A General Method for Encapsulating Macromolecules in Lipid Nanoparticle Systems","volume":"119","author":"Leung","year":"2015","journal-title":"J. Phys. Chem. B"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.jconrel.2023.03.039","article-title":"Continuous freeze-drying of messenger RNA lipid nanoparticles enables storage at higher temperatures","volume":"357","author":"Meulewaeter","year":"2023","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.jconrel.2015.06.028","article-title":"Multicomponent mannose-containing liposomes efficiently deliver RNA in murine immature dendritic cells and provide productive anti-tumour response in murine melanoma model","volume":"213","author":"Markov","year":"2015","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1089\/10430340360535751","article-title":"Stability of mRNA\/cationic lipid lipoplexes in human and rat cerebrospinal fluid: Methods and evidence for nonviral mRNA gene delivery to the central nervous system","volume":"14","author":"Anderson","year":"2003","journal-title":"Hum. Gene Ther."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1211\/002235702928","article-title":"Enhancement of polyethylene glycol (PEG)-modified cationic liposome-mediated gene deliveries: Effects on serum stability and transfection efficiency","volume":"55","author":"Kim","year":"2003","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"848","DOI":"10.3390\/pharmaceutics3040848","article-title":"Exploring the correlation between lipid packaging in lipoplexes and their transfection efficacy","volume":"3","author":"Moghaddam","year":"2011","journal-title":"Pharmaceutics"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"e2310491120","DOI":"10.1073\/pnas.2310491120","article-title":"pH-dependent structural transitions in cationic ionizable lipid mesophases are critical for lipid nanoparticle function","volume":"120","author":"Philipp","year":"2023","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"3631","DOI":"10.1002\/j.1460-2075.1994.tb06670.x","article-title":"Folding of firefly luciferase during translation in a cell-free system","volume":"13","author":"Kolb","year":"1994","journal-title":"EMBO J."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/j.jconrel.2015.08.007","article-title":"Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes","volume":"217","author":"Pardi","year":"2015","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_87","doi-asserted-by":"crossref","unstructured":"Zhang, W., Pfeifle, A., Lansdell, C., Frahm, G., Cecillon, J., Tamming, L., Gravel, C., Gao, J., Thulasi Raman, S.N., and Wang, L. (2023). The Expression Kinetics and Immunogenicity of Lipid Nanoparticles Delivering Plasmid DNA and mRNA in Mice. Vaccines, 11.","DOI":"10.3390\/vaccines11101580"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"e326","DOI":"10.1038\/mtna.2016.38","article-title":"Particle-mediated Intravenous Delivery of Antigen mRNA Results in Strong Antigen-specific T-cell Responses Despite the Induction of Type I Interferon","volume":"5","author":"Broos","year":"2016","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.jconrel.2012.12.029","article-title":"Transfection efficiency and transgene expression kinetics of mRNA delivered in naked and nanoparticle format","volume":"166","author":"Phua","year":"2013","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.jconrel.2015.08.051","article-title":"N(1)-methylpseudouridine-incorporated mRNA outperforms pseudouridine-incorporated mRNA by providing enhanced protein expression and reduced immunogenicity in mammalian cell lines and mice","volume":"217","author":"Andries","year":"2015","journal-title":"J. Control. Release Off. J. Control. 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