{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T19:19:56Z","timestamp":1770751196779,"version":"3.50.0"},"reference-count":82,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T00:00:00Z","timestamp":1770681600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Splicing defects represent a significant class of human genetic disorders, yet strategies to directly correct aberrant splice-site recognition remain limited. The small nuclear RNA (snRNA) U1 plays a critical role in pre-messenger RNA splicing by base-pairing with the conserved 5\u2032 splice-site \u2018GU\u2019 dinucleotide. Disruption of this interaction can lead to abnormal splicing or frameshift mutations, contributing to disease pathology. Extracellular vesicles (EVs) can transport small molecules to cells for therapeutic applications. Here, U1 snRNA-overexpressing HEK293T cells were used to generate approximately 120 nm-diameter U1 snRNA-enriched EVs, whose purity and content were confirmed by exosomal marker Western blots and reverse transcription\u2013quantitative PCR. When HeLa cells expressing a \u03b2-globin minigene bearing a \u03b2-thalassaemia-like 5\u2032 splice-site mutation were treated with U1-snRNA-enriched EVs, they corrected up to sixty percent of normal exon\u2013intron junction recognition in a dose-dependent manner. Recovery was abolished by heat or RNase treatment, suggesting that intact vesicular RNA cargo was essential for activity. These findings provide the first demonstration that EVs can transport spliceosomal snRNAs capable of reconstituting splice-site recognition in recipient cells and introduce a novel class of RNA-based therapeutics that exploit the natural cargo-shuttling capacity of EVs to correct splicing defects associated with genetic disease.<\/jats:p>","DOI":"10.3390\/biom16020278","type":"journal-article","created":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T12:07:06Z","timestamp":1770725226000},"page":"278","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Extracellular Vesicle-Mediated U1 snRNA Delivery Restores Aberrant Pre-mRNA Splicing in Human Cells"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9357-4975","authenticated-orcid":false,"given":"Hatice","family":"Esenkaya","sequence":"first","affiliation":[{"name":"Department of Cell and Molecular Biology, Karolinska Institutet, 17 177 Stockholm, Sweden"},{"name":"School of Life Sciences, Molecular Biology and Genetics, Kilis 7 Aralik University, 79000 Kilis, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Muhammet","family":"Karaman","sequence":"additional","affiliation":[{"name":"Department of Biology, Faculty of Science, Dokuz Eylul University, 35390 Izmir, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7434-6632","authenticated-orcid":false,"given":"Joe","family":"Bryant","sequence":"additional","affiliation":[{"name":"School of Biosciences, Faculty of Health and Medical Sciences, University of Surrey, Guildford GU2 7XH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1413","DOI":"10.1038\/ng.259","article-title":"Deep surveying of alternative splicing complexity in the human transcriptome by high-throughput sequencing","volume":"40","author":"Pan","year":"2008","journal-title":"Nat. Genet."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"387","DOI":"10.3233\/ISB-00142","article-title":"Distributions of exons and introns in the human genome","volume":"4","author":"Sakharkar","year":"2004","journal-title":"In Silico Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1016\/j.cell.2017.04.033","article-title":"An Atomic Structure of the Human Spliceosome","volume":"169","author":"Zhang","year":"2017","journal-title":"Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"9807","DOI":"10.1021\/acs.chemrev.5c00374","article-title":"Decoding Spliceosome Dynamics through Computation and Experiment","volume":"125","author":"Fica","year":"2025","journal-title":"Chem. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"102358","DOI":"10.1016\/j.gde.2025.102358","article-title":"Spliceosome complex and neurodevelopmental disorders","volume":"93","author":"Deutsch","year":"2025","journal-title":"Curr. Opin. Genet. Dev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1177\/13872877251359633","article-title":"Alternative splicing in Alzheimer\u2019s disease: Mechanisms, therapeutic implications, and 3D modeling approaches","volume":"107","author":"Pasteris","year":"2025","journal-title":"J. Alzheimer\u2019s Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e70026","DOI":"10.1002\/wrna.70026","article-title":"Role of the U1 snRNP Complex in Human Health and Disease","volume":"16","author":"Soares","year":"2025","journal-title":"WIREs RNA"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1038\/s41576-018-0028-8","article-title":"Refining the splice region","volume":"19","author":"Furlong","year":"2018","journal-title":"Nat. Rev. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1038\/283220a0","article-title":"Are snRNPs involved in splicing?","volume":"283","author":"Lerner","year":"1980","journal-title":"Nature"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1007\/BF00210743","article-title":"The mutational spectrum of single base-pair substitutions in mRNA splice junctions of human genes: Causes and consequences","volume":"90","author":"Krawczak","year":"1992","journal-title":"Hum. Genet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1002\/humu.20400","article-title":"Single base-pair substitutions in exon-intron junctions of human genes: Nature, distribution, and consequences for mRNA splicing","volume":"28","author":"Krawczak","year":"2007","journal-title":"Hum. Mutat."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1038\/302591a0","article-title":"Specific transcription and RNA splicing defects in five cloned \u03b2-thalassaemia genes","volume":"302","author":"Treisman","year":"1983","journal-title":"Nature"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1089\/hum.2023.202","article-title":"Gene Editing of the Endogenous Cryptic 3\u2032 Splice Site Corrects the RNA Splicing Defect in the \u03b2-Thalassemia Mouse Model","volume":"35","author":"Lu","year":"2024","journal-title":"Hum. Gene Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1038\/nrg.2015.3","article-title":"RNA mis-splicing in disease","volume":"17","author":"Scotti","year":"2016","journal-title":"Nat. Rev. Genet."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1038\/s41588-025-02184-4","article-title":"Dominant variants in major spliceosome U4 and U5 small nuclear RNA genes cause neurodevelopmental disorders through splicing disruption","volume":"57","author":"Nava","year":"2025","journal-title":"Nat. Genet."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e1532","DOI":"10.1002\/wrna.1532","article-title":"Splicing and neurodegeneration: Insights and mechanisms","volume":"10","author":"Nik","year":"2019","journal-title":"WIREs RNA"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Ottesen, E.W., Singh, N.N., Seo, J., and Singh, R.N. (2024). U1 snRNA interactions with deep intronic sequences regulate splicing of multiple exons of spinal muscular atrophy genes. Front. Neurosci., 18.","DOI":"10.3389\/fnins.2024.1412893"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e1476","DOI":"10.1002\/wrna.1476","article-title":"Alternative-splicing defects in cancer: Splicing regulators and their downstream targets, guiding the way to novel cancer therapeutics","volume":"9","author":"Urbanski","year":"2018","journal-title":"WIREs RNA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1038\/s41568-022-00541-7","article-title":"RNA splicing dysregulation and the hallmarks of cancer","volume":"23","author":"Bradley","year":"2023","journal-title":"Nat. Rev. Cancer"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1186\/s13046-025-03279-w","article-title":"Targeting RNA splicing modulation: New perspectives for anticancer strategy?","volume":"44","author":"Lv","year":"2025","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6549","DOI":"10.1093\/nar\/gkw533","article-title":"Splice-switching antisense oligonucleotides as therapeutic drugs","volume":"44","author":"Havens","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1816","DOI":"10.1038\/s12276-024-01292-1","article-title":"Tailored antisense oligonucleotides designed to correct aberrant splicing reveal actionable groups of mutations for rare genetic disorders","volume":"56","author":"Wai","year":"2024","journal-title":"Exp. Mol. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1007\/s40259-024-00644-7","article-title":"Splice-Modulating Antisense Oligonucleotides as Therapeutics for Inherited Metabolic Diseases","volume":"38","author":"Chen","year":"2024","journal-title":"BioDrugs"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1723","DOI":"10.1056\/NEJMoa1702752","article-title":"Nusinersen versus Sham Control in Infantile-Onset Spinal Muscular Atrophy","volume":"377","author":"Finkel","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1007\/s00415-025-12984-7","article-title":"Long-term CSF responses in adult patients with spinal muscular atrophy type 2 or 3 on treatment with nusinersen","volume":"272","author":"Cebulla","year":"2025","journal-title":"J. Neurol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1186\/s13073-025-01477-x","article-title":"Systematic analysis of genetic and phenotypic characteristics reveals antisense oligonucleotide therapy potential for one-third of neurodevelopmental disorders","volume":"17","author":"Wijnant","year":"2025","journal-title":"Genome Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"100182","DOI":"10.1016\/j.crtox.2024.100182","article-title":"Comprehensive review of adverse reactions and toxicology in ASO-based therapies for Duchenne Muscular Dystrophy: From FDA-approved drugs to peptide-conjugated ASO","volume":"7","author":"Kohut","year":"2024","journal-title":"Curr. Res. Toxicol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Raguraman, P., Balachandran, A.A., Chen, S., Diermeier, S.D., and Veedu, R.N. (2021). Antisense Oligonucleotide-Mediated Splice Switching: Potential Therapeutic Approach for Cancer Mitigation. Cancers, 13.","DOI":"10.3390\/cancers13215555"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Gagliardi, M., and Ashizawa, A.T. (2021). The Challenges and Strategies of Antisense Oligonucleotide Drug Delivery. Biomedicines, 9.","DOI":"10.3390\/biomedicines9040433"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1038\/s43856-023-00419-1","article-title":"Possibilities and limitations of antisense oligonucleotide therapies for the treatment of monogenic disorders","volume":"4","author":"Lauffer","year":"2024","journal-title":"Commun. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e13243","DOI":"10.15252\/emmm.202013243","article-title":"Delivery of oligonucleotide-based therapeutics: Challenges and opportunities","volume":"13","author":"Hammond","year":"2021","journal-title":"EMBO Mol. Med."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Qiu, J., Wu, L., Qu, R., Jiang, T., Bai, J., Sheng, L., Feng, P., and Sun, J. (2022). History of development of the life-saving drug \u201cNusinersen\u201d in spinal muscular atrophy. Front. Cell. Neurosci., 16.","DOI":"10.3389\/fncel.2022.942976"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1016\/0092-8674(86)90064-4","article-title":"A compensatory base change in U1 snRNA suppresses a 5\u2032 splice site mutation","volume":"46","author":"Zhuang","year":"1986","journal-title":"Cell"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1016\/j.ymeth.2022.06.008","article-title":"Engineered U1 snRNAs to modulate alternatively spliced exons","volume":"205","author":"Hatch","year":"2022","journal-title":"Methods"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2389","DOI":"10.1093\/hmg\/dds045","article-title":"An exon-specific U1 small nuclear RNA (snRNA) strategy to correct splicing defects","volume":"21","author":"Pinotti","year":"2012","journal-title":"Hum. Mol. Genet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.omtn.2019.08.014","article-title":"Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation","volume":"18","author":"Breuel","year":"2019","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1186\/s10020-025-01090-z","article-title":"Rescue of common and rare exon 2 skipping variants of the GAA gene using modified U1 snRNA","volume":"31","author":"Peruzzo","year":"2025","journal-title":"Mol. Med."},{"key":"ref_38","first-page":"7760556","article-title":"Characterization and Engineered U1 snRNA Rescue of Splicing Variants in a Turkish Neurodevelopmental Disease Cohort","volume":"2024","author":"Stuani","year":"2024","journal-title":"Hum. Mutat."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Gon\u00e7alves, M., Santos, J.I., Coutinho, M.F., Matos, L., and Alves, S. (2023). Development of Engineered-U1 snRNA Therapies: Current Status. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241914617"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1002\/humu.21509","article-title":"U1 snRNA-mediated gene therapeutic correction of splice defects caused by an exceptionally mild BBS mutation","volume":"32","author":"Schmid","year":"2011","journal-title":"Hum. Mutat."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1529","DOI":"10.1038\/s41375-018-0106-0","article-title":"Clinical use of lentiviral vectors","volume":"32","author":"Milone","year":"2018","journal-title":"Leukemia"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Dong, W., and Kantor, B. (2021). Lentiviral Vectors for Delivery of Gene-Editing Systems Based on CRISPR\/Cas: Current State and Perspectives. Viruses, 13.","DOI":"10.3390\/v13071288"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"654","DOI":"10.1038\/ncb1596","article-title":"Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells","volume":"9","author":"Valadi","year":"2007","journal-title":"Nat. Cell Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines","volume":"7","author":"Witwer","year":"2018","journal-title":"J. Extracell. Vesicles"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"eaau6977","DOI":"10.1126\/science.aau6977","article-title":"The biology, function, and biomedical applications of exosomes","volume":"367","author":"Kalluri","year":"2020","journal-title":"Science"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1038\/nrd3978","article-title":"Extracellular vesicles: Biology and emerging therapeutic opportunities","volume":"12","author":"Breakefield","year":"2013","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Andreu, Z., and Yanez-Mo, M. (2014). Tetraspanins in Extracellular Vesicle Formation and Function. Front. Immunol., 5.","DOI":"10.3389\/fimmu.2014.00442"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"4028","DOI":"10.1038\/s41467-025-59377-y","article-title":"Engineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors","volume":"16","author":"Liang","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Goo, J., Lee, Y., Lee, J., Kim, I.S., and Jeong, C. (2024). Extracellular Vesicles in Therapeutics: A Comprehensive Review on Applications, Challenges, and Clinical Progress. Pharmaceutics, 16.","DOI":"10.3390\/pharmaceutics16030311"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"4895","DOI":"10.1113\/JP282047","article-title":"Message in a bubble: The translational potential of extracellular vesicles","volume":"601","author":"Alqurashi","year":"2023","journal-title":"J. Physiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1038\/s44222-025-00277-7","article-title":"Extracellular vesicles for the delivery of gene therapy","volume":"3","author":"Obuchi","year":"2025","journal-title":"Nat. Rev. Bioeng."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"e12458","DOI":"10.1002\/jev2.12458","article-title":"A systematic review and meta-analysis of clinical trials assessing safety and efficacy of human extracellular vesicle-based therapy","volume":"13","author":"Derdelinckx","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"e04986","DOI":"10.7554\/eLife.04986","article-title":"Crystal structure of human U1 snRNP, a small nuclear ribonucleoprotein particle, reveals the mechanism of 5\u2032 splice site recognition","volume":"4","author":"Kondo","year":"2015","journal-title":"eLife"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"108958","DOI":"10.1016\/j.pharmthera.2025.108958","article-title":"RNA-based drugs: Current, imminent and possible therapeutic applications","volume":"277","author":"Torrisi","year":"2026","journal-title":"Pharmacol. Ther."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"118495","DOI":"10.1016\/j.ejmech.2025.118495","article-title":"Unveiling RNA ligands: Harnessing NMR for hit generation","volume":"304","author":"Gaspar","year":"2026","journal-title":"Eur. J. Med. Chem."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"2980","DOI":"10.1038\/ncomms3980","article-title":"Sumoylated hnRNPA2B1 controls the sorting of miRNAs into exosomes through binding to specific motifs","volume":"4","author":"Mittelbrunn","year":"2013","journal-title":"Nat. Commun."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1016\/j.celrep.2016.09.031","article-title":"The RNA-Binding Protein SYNCRIP Is a Component of the Hepatocyte Exosomal Machinery Controlling MicroRNA Sorting","volume":"17","author":"Santangelo","year":"2016","journal-title":"Cell Rep."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"105834","DOI":"10.1016\/j.ebiom.2025.105834","article-title":"Viral and non-viral vectors in gene therapy: Current state and clinical perspectives","volume":"118","author":"Geng","year":"2025","journal-title":"eBioMedicine"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1007\/s12033-024-01070-4","article-title":"The Future of Gene Therapy: A Review of In Vivo and Ex Vivo Delivery Methods for Genome Editing-Based Therapies","volume":"67","author":"Volodina","year":"2025","journal-title":"Mol. Biotechnol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.1016\/j.ymthe.2023.01.021","article-title":"Extracellular vesicles: The next generation in gene therapy delivery","volume":"31","author":"Cecchin","year":"2023","journal-title":"Mol. Ther."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.1038\/s41467-018-03733-8","article-title":"Designer exosomes produced by implanted cells intracerebrally deliver therapeutic cargo for Parkinson\u2019s disease treatment","volume":"9","author":"Kojima","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"24641","DOI":"10.3402\/jev.v3.24641","article-title":"Routes and mechanisms of extracellular vesicle uptake","volume":"3","author":"Mulcahy","year":"2014","journal-title":"J. Extracell. Vesicles"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1038\/nrm3742","article-title":"A day in the life of the spliceosome","volume":"15","author":"Matera","year":"2014","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"4444","DOI":"10.1021\/acsnano.9b10033","article-title":"Endocytosis of Extracellular Vesicles and Release of Their Cargo from Endosomes","volume":"14","author":"Joshi","year":"2020","journal-title":"ACS Nano"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"7029","DOI":"10.1038\/ncomms8029","article-title":"Visualization and tracking of tumour extracellular vesicle delivery and RNA translation using multiplexed reporters","volume":"6","author":"Lai","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"828","DOI":"10.1038\/s41573-025-01237-x","article-title":"RNA chemistry and therapeutics","volume":"24","author":"Wang","year":"2025","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1111\/jth.12471","article-title":"An engineered U1 small nuclear RNA rescues splicing-defective coagulation F7 gene expression in mice","volume":"12","author":"Balestra","year":"2014","journal-title":"J. Thromb. Haemost."},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Dong, X., Huang, Y., Yi, T., Hu, C., Gao, Q., Chen, Y., Zhang, J., Chen, J., Liu, L., and Meng, R. (2022). Intrapleural infusion of tumor cell-derived microparticles packaging methotrexate or saline combined with pemetrexed-cisplatin chemotherapy for the treatment of malignant pleural effusion in advanced non-squamous non-small cell lung cancer: A double-blind, randomized, placebo-controlled study. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.1002938"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1186\/s13287-022-02900-5","article-title":"Nebulized exosomes derived from allogenic adipose tissue mesenchymal stromal cells in patients with severe COVID-19: A pilot study","volume":"13","author":"Zhu","year":"2022","journal-title":"Stem Cell Res. Ther."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"e15997","DOI":"10.15252\/emmm.202215997","article-title":"A novel platform for attenuating immune hyperactivity using EXO-CD24 in COVID-19 and beyond","volume":"14","author":"Shapira","year":"2022","journal-title":"EMBO Mol. Med."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"890","DOI":"10.1212\/WNL.0000000000002445","article-title":"Results from a phase 1 study of nusinersen in children with spinal muscular atrophy","volume":"86","author":"Chiriboga","year":"2016","journal-title":"Neurology"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"e202000889","DOI":"10.26508\/lsa.202000889","article-title":"Short-duration splice promoting compound enables a tunable mouse model of spinal muscular atrophy","volume":"4","author":"Rietz","year":"2021","journal-title":"Life Sci. Alliance"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.nmd.2018.01.003","article-title":"Correlation between SMA type and SMN2 copy number revisited: An analysis of 625 unrelated Spanish patients and a compilation of 2834 reported cases","volume":"28","author":"Calucho","year":"2018","journal-title":"Neuromuscul. Disord."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0268-960X(94)90002-7","article-title":"Genotype-phenotype correlations in \u03b2-thalassemias","volume":"8","author":"Cao","year":"1994","journal-title":"Blood Rev."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"12277","DOI":"10.1038\/ncomms12277","article-title":"Exosome engineering for efficient intracellular delivery of soluble proteins using optically reversible protein\u2013protein interaction module","volume":"7","author":"Yim","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.cbpa.2019.11.002","article-title":"High-throughput approaches to profile RNA-protein interactions","volume":"54","author":"Nechay","year":"2020","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"114021","DOI":"10.1016\/j.jconrel.2025.114021","article-title":"Recent advances and challenges of injectable hydrogels in drug delivery","volume":"385","author":"Guo","year":"2025","journal-title":"J. Control. Release"},{"key":"ref_78","doi-asserted-by":"crossref","unstructured":"Liu, B., and Chen, K. (2024). Advances in Hydrogel-Based Drug Delivery Systems. Gels, 10.","DOI":"10.3390\/gels10040262"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1038\/nbt.1807","article-title":"Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes","volume":"29","author":"Seow","year":"2011","journal-title":"Nat. Biotechnol."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"1597603","DOI":"10.1080\/20013078.2019.1597603","article-title":"High-fidelity detection and sorting of nanoscale vesicles in viral disease and cancer","volume":"8","author":"Musich","year":"2019","journal-title":"J. Extracell. Vesicles"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"e70094","DOI":"10.1002\/jev2.70094","article-title":"Engineering of CD63 Enables Selective Extracellular Vesicle Cargo Loading and Enhanced Payload Delivery","volume":"14","author":"Obuchi","year":"2025","journal-title":"J. Extracell. Vesicles"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"100031","DOI":"10.1016\/j.vesic.2023.100031","article-title":"CD63-snorkel tagging for isolation of exosomes","volume":"2","author":"Han","year":"2023","journal-title":"Extracell. 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