{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T21:02:24Z","timestamp":1791838944351,"version":"4.3.4"},"reference-count":24,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T00:00:00Z","timestamp":1771891200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003593","name":"National Council for Scientific and Technological Development","doi-asserted-by":"crossref","award":["470888\/2014-8"],"award-info":[{"award-number":["470888\/2014-8"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100003593","name":"National Council for Scientific and Technological Development","doi-asserted-by":"crossref","award":["141742\/2014-3"],"award-info":[{"award-number":["141742\/2014-3"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100003593","name":"National Council for Scientific and Technological Development","doi-asserted-by":"crossref","award":["403587\/2021-2"],"award-info":[{"award-number":["403587\/2021-2"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Financiamento e Incentivo \u00e0 Pesquisa of Hospital de Cl\u00ednicas de Porto Alegre FIPE\/HCPA","award":["2019-0122"],"award-info":[{"award-number":["2019-0122"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Background\/Objectives: Mucopolysaccharidosis type I (MPS I) is a lysosomal storage disorder caused by \u03b1-L-iduronidase (IDUA) deficiency, leading to progressive glycosaminoglycan (GAG) accumulation and severe joint involvement. Gene editing represents a promising alternative to restore localized enzyme production. Therefore, this study aimed to evaluate the feasibility, efficacy, and safety of in situ genome editing through intra-articular administration of a nonviral CRISPR\/Cas9 system to increase localized IDUA expression in an MPS I mouse model. Methods: Cationic liposomes were formulated to deliver plasmids encoding the CRISPR\/Cas9 system targeted to the ROSA26 locus along with an IDUA donor sequence. In vitro assays were performed in fibroblast-like synoviocytes (FLSs) isolated from MPS I mice to assess cytotoxicity, gene editing efficiency, and IDUA activity. In vivo, MPS I mice received intra-articular injections in the knee joints, either as a single dose (short-term study) or monthly for three months (long-term study). IDUA activity, GAG levels, and genome editing efficiency were evaluated in joint tissues, synovial fluid, serum, and major organs. Results: Gene-edited FLS showed sustained IDUA activity for up to 30 days with low cytotoxicity. In vivo, intra-articular administration resulted in a significant increase in IDUA activity in joint tissue and synovial fluid without detectable systemic IDUA. Long-term treatment led to persistent joint-localized IDUA activity, significant reductions (&gt;50%) in GAG levels, and detectable genome editing in joint DNA. Conclusions: Intra-articular delivery of CRISPR\/Cas9 via cationic liposomes enables safe and effective localized genome editing, representing a promising strategy for treating joint manifestations of MPS I.<\/jats:p>","DOI":"10.3390\/pharmaceutics18030281","type":"journal-article","created":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T08:54:56Z","timestamp":1771923296000},"page":"281","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Liposomal CRISPR\/Cas9-Mediated Local Genome Editing for Joint Disease in Mucopolysaccharidosis Type I"],"prefix":"10.3390","volume":"18","author":[{"given":"Hallana Souza","family":"Santos","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio C\u00e9lulas, Tecidos e Genes do Hospital de Cl\u00ednicas de Porto Alegre, R. Ramiro Barcelos, 2350, Porto Alegre 90035-903, RS, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Gen\u00e9tica e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Gen\u00e9tica, Campus do Vale, Av. Bento Gon\u00e7alves, 9500, Porto Alegre 91501-970, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0554-0295","authenticated-orcid":false,"given":"Edina","family":"Poletto","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio C\u00e9lulas, Tecidos e Genes do Hospital de Cl\u00ednicas de Porto Alegre, R. Ramiro Barcelos, 2350, Porto Alegre 90035-903, RS, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Gen\u00e9tica e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Gen\u00e9tica, Campus do Vale, Av. Bento Gon\u00e7alves, 9500, Porto Alegre 91501-970, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luisa Natalia Pimentel","family":"Vera","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio C\u00e9lulas, Tecidos e Genes do Hospital de Cl\u00ednicas de Porto Alegre, R. Ramiro Barcelos, 2350, Porto Alegre 90035-903, RS, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Gen\u00e9tica e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Gen\u00e9tica, Campus do Vale, Av. Bento Gon\u00e7alves, 9500, Porto Alegre 91501-970, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mirian","family":"Farinon","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias M\u00e9dicas da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Medicina, Campus Sa\u00fade, Av. Ramiro Barcelos, 2400, Porto Alegre 90035-003, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9330-0154","authenticated-orcid":false,"given":"Francyne","family":"Kubaski","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Gen\u00e9tica e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Gen\u00e9tica, Campus do Vale, Av. Bento Gon\u00e7alves, 9500, Porto Alegre 91501-970, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8401-3332","authenticated-orcid":false,"given":"Paola Barcelos","family":"Carneiro","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio C\u00e9lulas, Tecidos e Genes do Hospital de Cl\u00ednicas de Porto Alegre, R. Ramiro Barcelos, 2350, Porto Alegre 90035-903, RS, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Gen\u00e9tica e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Gen\u00e9tica, Campus do Vale, Av. Bento Gon\u00e7alves, 9500, Porto Alegre 91501-970, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2820-2814","authenticated-orcid":false,"given":"Willian da Silva","family":"Carniel","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias Farmac\u00eauticas da Universidade Federal do Rio Grande do Sul (UFRGS), Faculdade de Farm\u00e1cia, Av. Ipiranga, 2752, Porto Alegre 90610-000, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9655-3686","authenticated-orcid":false,"given":"Roberto","family":"Giugliani","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Gen\u00e9tica e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Gen\u00e9tica, Campus do Vale, Av. Bento Gon\u00e7alves, 9500, Porto Alegre 91501-970, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4977-6662","authenticated-orcid":false,"given":"Ursula","family":"Matte","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio C\u00e9lulas, Tecidos e Genes do Hospital de Cl\u00ednicas de Porto Alegre, R. Ramiro Barcelos, 2350, Porto Alegre 90035-903, RS, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Gen\u00e9tica e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Gen\u00e9tica, Campus do Vale, Av. Bento Gon\u00e7alves, 9500, Porto Alegre 91501-970, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helder Ferreira","family":"Teixeira","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias Farmac\u00eauticas da Universidade Federal do Rio Grande do Sul (UFRGS), Faculdade de Farm\u00e1cia, Av. Ipiranga, 2752, Porto Alegre 90610-000, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4189-0436","authenticated-orcid":false,"given":"Roselena Silvestri","family":"Schuh","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio C\u00e9lulas, Tecidos e Genes do Hospital de Cl\u00ednicas de Porto Alegre, R. Ramiro Barcelos, 2350, Porto Alegre 90035-903, RS, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias Farmac\u00eauticas da Universidade Federal do Rio Grande do Sul (UFRGS), Faculdade de Farm\u00e1cia, Av. Ipiranga, 2752, Porto Alegre 90610-000, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1598-5861","authenticated-orcid":false,"given":"Guilherme","family":"Baldo","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio C\u00e9lulas, Tecidos e Genes do Hospital de Cl\u00ednicas de Porto Alegre, R. Ramiro Barcelos, 2350, Porto Alegre 90035-903, RS, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Gen\u00e9tica e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Gen\u00e9tica, Campus do Vale, Av. Bento Gon\u00e7alves, 9500, Porto Alegre 91501-970, RS, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Fisiologia da Universidade Federal do Rio Grande do Sul (UFRGS), Instituto de Ci\u00eancias B\u00e1sicas da Sa\u00fade, R. Sarmento Leite, 500, Porto Alegre 90035-190, RS, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/j.rchipe.2015.10.004","article-title":"Mucopolisacaridosis: Caracter\u00edsticas cl\u00ednicas, diagn\u00f3stico y de manejo [Mucopolysaccharidosis: Clinical features, diagnosis and management]","volume":"87","year":"2016","journal-title":"Rev. Chil. Pediatr."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e1228","DOI":"10.1542\/peds.2008-0999","article-title":"Multidisciplinary Management of Hunter Syndrome","volume":"124","author":"Muenzer","year":"2009","journal-title":"Pediatrics"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1813","DOI":"10.1016\/j.joca.2013.08.001","article-title":"Osteoimmunology in Mucopolysaccharidoses Type I, II, VI and VII. Immunological Regulation of the Osteoarticular System in the Course Ofmetabolic Inflammation","volume":"21","author":"Jurecka","year":"2013","journal-title":"Osteoarthr. Cartil."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1542\/peds.2008-0416","article-title":"International Consensus Panel on Management and Treatment of Mucopolysaccharidosis I. Mucopolysaccharidosis I: Management and treatment guidelines","volume":"123","author":"Muenzer","year":"2009","journal-title":"Pediatrics"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.ijpharm.2018.06.049","article-title":"Intra-Articular Nonviral Gene Therapy in Mucopolysaccharidosis I Mice","volume":"548","author":"Bidone","year":"2018","journal-title":"Int. J. Pharm."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1007\/s11095-018-2503-5","article-title":"Nasal Administration of Cationic Nanoemulsions as Nucleic Acids Delivery Systems Aiming at Mucopolysaccharidosis Type I Gene Therapy","volume":"35","author":"Schuh","year":"2018","journal-title":"Pharm. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.5582\/irdr.2020.01011","article-title":"A basic understanding of mucopolysaccharidosis: Incidence, clinical features, diagnosis, and management","volume":"9","author":"Zhou","year":"2020","journal-title":"Intractable Rare Dis. Res."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Poletto, E., Baldo, G., and Gomez-Ospina, N. (2020). Genome Editing for Mucopolysaccharidoses. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21020500"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1088","DOI":"10.1002\/jimd.12414","article-title":"Mucopolysaccharidoses type I gene therapy","volume":"44","author":"Hurt","year":"2021","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_10","first-page":"79","article-title":"The Role of Gene Therapy in Cartilage Repair","volume":"7","author":"Valentino","year":"2019","journal-title":"Arch. Bone Jt. Surg."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1089\/hum.2017.181","article-title":"Gene Delivery to Joints by Intra-Articular Injection","volume":"29","author":"Evans","year":"2018","journal-title":"Hum. Gene Ther."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.jconrel.2018.08.031","article-title":"In Vivo Genome Editing of Mucopolysaccharidosis I Mice Using the CRISPR\/Cas9 System","volume":"288","author":"Schuh","year":"2018","journal-title":"J. Control. Release"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1159\/000327532","article-title":"Recombinant Encapsulated Cells Overexpressing Alpha-L-Iduronidase Correct Enzyme Deficiency in Human Mucopolysaccharidosis Type I Cells","volume":"195","author":"Baldo","year":"2012","journal-title":"Cells Tissues Organs"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Armaka, M., Gkretsi, V., Kontoyiannis, D., and Kollias, G. (2009). A Standardized Protocol for the Isolation and Culture of Normal and Arthritogenic Murine Synovial Fibroblasts. Protoc. Exch.","DOI":"10.1038\/nprot.2009.102"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0022-1759(83)90303-4","article-title":"Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assays","volume":"65","author":"Mosmann","year":"1983","journal-title":"J. Immunol. Methods"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1111\/j.1365-2249.2004.02499.x","article-title":"Joint Cytokine Quantification in Two Rodent Arthritis Models: Kinetics of Expression, Correlation of MRNA and Protein Levels and Response to Prednisolone Treatment","volume":"137","author":"Rioja","year":"2004","journal-title":"Clin. Exp. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/S0021-9258(19)52451-6","article-title":"Protein Measurement with the Folin Phenol Reagent","volume":"193","author":"Lowry","year":"1951","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1883","DOI":"10.1097\/01.ju.0000158448.56888.09","article-title":"Renal Medullary Carcinoma and ABL Gene Amplification","volume":"173","author":"Simpson","year":"2005","journal-title":"J. Urol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e3410","DOI":"10.1002\/jgm.3410","article-title":"Brain and Visceral Gene Editing of Mucopolysaccharidosis I Mice by Nasal Delivery of the CRISPR\/Cas9 System","volume":"24","author":"Vera","year":"2022","journal-title":"J. Gene Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"247","DOI":"10.2144\/000112913","article-title":"Real-Time PCR to Determine Transgene Copy Number and to Quantitate the Biolocalization of Adoptively Transferred Cells from EGFP-Transgenic Mice","volume":"45","author":"Joshi","year":"2008","journal-title":"Biotechniques"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.ejpb.2017.10.017","article-title":"Gene Editing of MPS I Human Fibroblasts by Co-Delivery of a CRISPR\/Cas9 Plasmid and a Donor Oligonucleotide Using Nanoemulsions as Nonviral Carriers","volume":"122","author":"Schuh","year":"2018","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1038\/labinvest.3780345","article-title":"Articular Chondrocytes from Animals with a Dermatan Sulfate Storage Disease Undergo a High Rate of Apoptosis and Release Nitric Oxide and Inflammatory Cytokines: A Possible Mechanism Underlying Degenerative Joint Disease in the Mucopolysaccharidoses","volume":"81","author":"Simonaro","year":"2001","journal-title":"Lab. Investig."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"100132","DOI":"10.1016\/j.onano.2023.100132","article-title":"Recent advances in using liposomes for delivery of nucleic acid-based therapeutics","volume":"11","author":"Nsairat","year":"2023","journal-title":"OpenNano"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1016\/j.ymgme.2014.05.015","article-title":"Intra-Articular Enzyme Replacement Therapy with RhIDUA Is Safe, Well-Tolerated, and Reduces Articular GAG Storage in the Canine Model of Mucopolysaccharidosis Type I","volume":"112","author":"Wang","year":"2014","journal-title":"Mol. Genet. Metab."}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/3\/281\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T05:20:24Z","timestamp":1772083224000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/3\/281"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,24]]},"references-count":24,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["pharmaceutics18030281"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics18030281","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,24]]}}}