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Under physiological conditions, GM2 ganglioside is catabolized by the \u03b2-hexosaminidase A in a GM2 activator protein-dependent mechanism. In contrast, uncharged substrates such as globosides and some glycosaminoglycans can be hydrolyzed by the \u03b2-hexosaminidase B. Monogenic mutations on HEXA, HEXB, or GM2A genes arise in the Tay\u2013Sachs (TSD), Sandhoff (SD), and AB variant diseases, respectively. In this work, we validated a CRISPR\/Cas9-based gene editing strategy that relies on a Cas9 nickase (nCas9) as a potential approach for treating GM2 gangliosidoses using in vitro models for TSD and SD. The nCas9 contains a mutation in the catalytic RuvC domain but maintains the active HNH domain, which reduces potential off-target effects. Liposomes (LPs)- and novel magnetoliposomes (MLPs)-based vectors were used to deliver the CRISPR\/nCas9 system. When LPs were used as a vector, positive outcomes were observed for the \u03b2-hexosaminidase activity, glycosaminoglycans levels, lysosome mass, and oxidative stress. In the case of MLPs, a high cytocompatibility and transfection ratio was observed, with a slight increase in the \u03b2-hexosaminidase activity and significant oxidative stress recovery in both TSD and SD cells. These results show the remarkable potential of CRISPR\/nCas9 as a new alternative for treating GM2 gangliosidoses, as well as the superior performance of non-viral vectors in enhancing the potency of this therapeutic approach.<\/jats:p>","DOI":"10.3390\/ijms231810672","type":"journal-article","created":{"date-parts":[[2022,9,14]],"date-time":"2022-09-14T20:50:45Z","timestamp":1663188645000},"page":"10672","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["CRISPR\/nCas9-Based Genome Editing on GM2 Gangliosidoses Fibroblasts via Non-Viral Vectors"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5956-1986","authenticated-orcid":false,"given":"Andr\u00e9s Felipe","family":"Leal","sequence":"first","affiliation":[{"name":"Institute for the Study of Inborn Errors of Metabolism, Faculty of Science, Pontificia Universidad Javeriana, Bogot\u00e1 110231, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0916-3909","authenticated-orcid":false,"given":"Javier","family":"Cifuentes","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Universidad de los Andes, Bogot\u00e1 111711, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2053-1303","authenticated-orcid":false,"given":"Valentina","family":"Quezada","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Universidad de los Andes, Bogot\u00e1 111711, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9791-2716","authenticated-orcid":false,"given":"Eliana","family":"Benincore-Fl\u00f3rez","sequence":"additional","affiliation":[{"name":"Institute for the Study of Inborn Errors of Metabolism, Faculty of Science, Pontificia Universidad Javeriana, Bogot\u00e1 110231, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7790-7546","authenticated-orcid":false,"given":"Juan Carlos","family":"Cruz","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Universidad de los Andes, Bogot\u00e1 111711, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7251-5298","authenticated-orcid":false,"given":"Luis Humberto","family":"Reyes","sequence":"additional","affiliation":[{"name":"Grupo de Dise\u00f1o de Productos y Procesos (GDPP), Department of Chemical and Food Engineering, Universidad de los Andes, Bogot\u00e1 111711, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9670-6435","authenticated-orcid":false,"given":"Angela Johana","family":"Espejo-Mojica","sequence":"additional","affiliation":[{"name":"Institute for the Study of Inborn Errors of Metabolism, Faculty of Science, Pontificia Universidad Javeriana, Bogot\u00e1 110231, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6484-1173","authenticated-orcid":false,"given":"Carlos Javier","family":"Alm\u00e9ciga-D\u00edaz","sequence":"additional","affiliation":[{"name":"Institute for the Study of Inborn Errors of Metabolism, Faculty of Science, Pontificia Universidad Javeriana, Bogot\u00e1 110231, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"100900","DOI":"10.1016\/j.jbior.2022.100900","article-title":"Sphingolipids and their role in health and disease in the central nervous system","volume":"85","author":"Leal","year":"2022","journal-title":"Adv. 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