{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T06:16:12Z","timestamp":1783577772344,"version":"3.55.0"},"reference-count":75,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2019,11,24]],"date-time":"2019-11-24T00:00:00Z","timestamp":1574553600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/BIM-MEC\/4762\/2014"],"award-info":[{"award-number":["PTDC\/BIM-MEC\/4762\/2014"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Sphingolipidoses are inherited genetic diseases characterized by the accumulation of glycosphingolipids. Sphingolipidoses (SP), which usually involve the loss of sphingolipid hydrolase function, are of lysosomal origin, and represent an important group of rare diseases among lysosomal storage disorders. Initial treatments consisted of enzyme replacement therapy, but, in recent decades, various therapeutic approaches have been developed. However, these commonly used treatments for SP fail to be fully effective and do not penetrate the blood\u2013brain barrier. New approaches, such as genome editing, have great potential for both the treatment and study of sphingolipidoses. Here, we review the most recent advances in the treatment and modelling of SP through the application of CRISPR-Cas9 genome editing. CRISPR-Cas9 is currently the most widely used method for genome editing. This technique is versatile; it can be used for altering the regulation of genes involved in sphingolipid degradation and synthesis pathways, interrogating gene function, generating knock out models, or knocking in mutations. CRISPR-Cas9 genome editing is being used as an approach to disease treatment, but more frequently it is utilized to create models of disease. New CRISPR-Cas9-based tools of gene editing with diminished off-targeting effects are evolving and seem to be more promising for the correction of individual mutations. Emerging Prime results and CRISPR-Cas9 difficulties are also discussed.<\/jats:p>","DOI":"10.3390\/ijms20235897","type":"journal-article","created":{"date-parts":[[2019,11,25]],"date-time":"2019-11-25T03:10:00Z","timestamp":1574651400000},"page":"5897","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Advances in Sphingolipidoses: CRISPR-Cas9 Editing as an Option for Modelling and Therapy"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9848-4805","authenticated-orcid":false,"given":"Renato","family":"Santos","sequence":"first","affiliation":[{"name":"Human Genetics Department, Unit of Research and Development, Nacional Health Institute Doctor Ricardo Jorge, 4000-055 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3478-2122","authenticated-orcid":false,"given":"Olga","family":"Amaral","sequence":"additional","affiliation":[{"name":"Human Genetics Department, Unit of Research and Development, Nacional Health Institute Doctor Ricardo Jorge, 4000-055 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,11,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1002\/path.3021","article-title":"The cellular pathology of lysosomal diseases","volume":"226","author":"Cox","year":"2012","journal-title":"J. Pathol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1038\/s41572-018-0025-4","article-title":"Lysosomal storage diseases","volume":"4","author":"Platt","year":"2018","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1136\/jcp.27.Suppl_8.94","article-title":"Sphingolipidoses","volume":"8","author":"Sandhoff","year":"1974","journal-title":"J. Clin. Pathol. Suppl."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1038\/nature13476","article-title":"Sphingolipid lysosomal storage disorders","volume":"510","author":"Platt","year":"2014","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1685","DOI":"10.4155\/fmc-2017-0065","article-title":"Recent advances and novel treatments for sphingolipidoses","volume":"9","author":"Arenz","year":"2017","journal-title":"Future Med. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s13024-019-0336-2","article-title":"Glucocerebrosidase and its relevance to Parkinson disease","volume":"14","author":"Do","year":"2019","journal-title":"Mol. Neurodegener."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1002\/mds.27631","article-title":"Emerging links between pediatric lysosomal storage diseases and adult parkinsonism","volume":"34","author":"Ysselstein","year":"2019","journal-title":"Mov Disord."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1644","DOI":"10.1038\/s41590-019-0504-0","article-title":"Control of CD1d-restricted antigen presentation and inflammation by sphingomyelin","volume":"20","author":"Melum","year":"2019","journal-title":"Nat. Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1126\/science.1174447","article-title":"A gene network regulating lysosomal biogenesis and function","volume":"325","author":"Sardiello","year":"2009","journal-title":"Science"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3852","DOI":"10.1093\/hmg\/ddr306","article-title":"Characterization of the CLEAR network reveals an integrated control of cellular clearance pathways","volume":"20","author":"Kang","year":"2011","journal-title":"Hum. Mol. Genet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1016\/j.tins.2019.05.010","article-title":"Can GBA1-Associated Parkinson Disease Be Modeled in the Mouse?","volume":"42","author":"Do","year":"2019","journal-title":"Trends Neurosci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1121","DOI":"10.1002\/humu.23056","article-title":"New Directions in Gaucher Disease","volume":"37","author":"Horowitz","year":"2016","journal-title":"Hum. Mutat."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1182","DOI":"10.1177\/1060028013500469","article-title":"Gaucher Disease and Its Treatment Options","volume":"47","author":"Bennett","year":"2013","journal-title":"Ann. Pharmacother."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1002\/ajh.24957","article-title":"GOS Study group.Demographics and patient characteristics of 1209 patients with Gaucher disease: Descriptive analysis from the Gaucher Outcome Survey (GOS)","volume":"93","author":"Zimran","year":"2018","journal-title":"Am. J. Hematol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1398","DOI":"10.1002\/humu.22124","article-title":"Gaucher disease paradigm: From ERAD to comorbidity","volume":"33","author":"Horowitz","year":"2012","journal-title":"Hum. Mutat."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1768","DOI":"10.1093\/hmg\/ddz005","article-title":"The effect of mutant GBA1 on accumulation and aggregation of \u03b1-synuclein","volume":"28","author":"Maor","year":"2019","journal-title":"Hum. Mol. Genet."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2706","DOI":"10.1093\/brain\/awx221","article-title":"The L444P Gba1 mutation enhances alpha-synuclein induced loss of nigral dopaminergic neurons in mice","volume":"140","author":"Wegrzynowicz","year":"2017","journal-title":"Brain"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Duro, G., Zizzo, C., Cammarata, G., Burlina, A., Burlina, A., Polo, G., Scalia, S., Oliveri, R., Sciarrino, S., and Francofonte, D. (2018). Mutations in the GLA Gene and LysoGb3: Is It Really Anderson-Fabry Disease?. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19123726"},{"key":"ref_19","first-page":"100454","article-title":"The effect of enzyme replacement therapy on clinical outcomes in male patients with Fabry disease: A systematic literature review by a European panel of experts","volume":"19","author":"Germain","year":"2019","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_20","unstructured":"Mehta, A., Beck, M., and Sunder-Plassmann, G. (2006). Chapter 19 Natural history of Fabry disease. Fabry Dis Ease: Perspectives from 5 Years of FOS, Oxford PharmaGenesis."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Waldek, S., and Feriozzi, S. (2014). Fabry nephropathy: A review\u2014How can we optimize the management of Fabry nephropathy?. BMC Nephrol., 15.","DOI":"10.1186\/1471-2369-15-72"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1186\/s13023-018-0886-3","article-title":"Neural stem cells for disease modeling and evaluation of therapeutics for Tay-Sachs disease","volume":"13","author":"Vu","year":"2018","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_23","first-page":"16.1","article-title":"Diagnosing Lysosomal Storage Disorders: The GM2 Gangliosidoses","volume":"83","author":"Hall","year":"2014","journal-title":"Curr. Protoc. Hum. Genet."},{"key":"ref_24","first-page":"55","article-title":"GM2-Gangliosidosis (Sandhoff and Tay Sachs disease): Diagnosis and Neuroimaging Findings (An Iranian Pediatric Case Series)","volume":"8","author":"Karimzadeh","year":"2014","journal-title":"Iran. J. Child Neurol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1663","DOI":"10.3389\/fphys.2018.01663","article-title":"New Approaches to Tay-Sachs Disease Therapy","volume":"9","author":"Solovyeva","year":"2018","journal-title":"Front. Physiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1016\/j.arcmed.2017.04.001","article-title":"Mutation Frequency of Three Neurodegenerative Lysosomal Storage Diseases: From Screening to Treatment?","volume":"48","author":"Duarte","year":"2017","journal-title":"Arch. Med. Res."},{"key":"ref_27","unstructured":"Adam, M.P., Ardinger, H.H., Pagon, R.A., Wallace, S.E., Bean, L.J.H., Stephens, K., and Amemiya, A. (1993). Hexosaminidase A Deficiency. GeneReviews, University of Washington."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1717","DOI":"10.1016\/B978-0-444-59565-2.00041-1","article-title":"Niemann-Pick diseases","volume":"113","author":"Vanier","year":"2013","journal-title":"Handb. Clin. Neurol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1186\/s13023-019-0996-6","article-title":"Disease characteristics, prognosis and miglustat treatment effects on disease progression in patients with Niemann-Pick disease Type C: An international, multicenter, retrospective chart review","volume":"14","author":"Pineda","year":"2019","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1086\/503750","article-title":"Imprinting at the SMPD1 Locus: Implications for Acid Sphingomyelinase\u2014Deficient Niemann-Pick Disease","volume":"78","author":"Simonaro","year":"2006","journal-title":"Am. J. Hum. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.ymgme.2016.12.008","article-title":"Types A and B Niemann-Pick disease","volume":"120","author":"Schuchman","year":"2017","journal-title":"Mol. Genet. Metab."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.gene.2014.09.046","article-title":"History, genetic, and recent advances on Krabbe disease","volume":"555","author":"Graziano","year":"2015","journal-title":"Gene"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"908","DOI":"10.1111\/tra.12404","article-title":"Molecular Mechanisms of Disease Pathogenesis Differ in Krabbe Disease Variants","volume":"17","author":"Spratley","year":"2016","journal-title":"Traffic"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1186\/s13023-018-0872-9","article-title":"A prospective natural history study of Krabbe disease in a patient cohort with onset between 6 months and 3 years of life","volume":"13","author":"Bascou","year":"2018","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_35","unstructured":"Scriver, C.R., Beaudet, A.L., Sly, W.S., and Valle, D. (2001). \u03b2-Galactosidase deficiency (\u03b2-galactosidosis) GM1 gangliosidosis and Morquio B disease. The Metabolic and Molecular Bases of Inherited Disease, McGraw-Hill."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1038\/jhg.2009.70","article-title":"Structural bases of GM1 gangliosidosis and Morquio B disease","volume":"54","author":"Morita","year":"2009","journal-title":"J. Hum. Genet."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"782","DOI":"10.1016\/j.bbadis.2011.03.018","article-title":"GM1 gangliosidosis and Morquio B disease: An update on genetic alterations and clinical findings","volume":"1812","author":"Caciotti","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1464","DOI":"10.1056\/NEJM199105233242104","article-title":"Replacement therapy for inherited enzyme deficiency\u2014Macrophage-targeted glucocerebrosidase for Gaucher\u2019s disease","volume":"324","author":"Barton","year":"1991","journal-title":"N. Engl. J. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1016\/j.tibtech.2013.04.004","article-title":"ZFN, TALEN and CRISPR\/Cas-based methods for genome engineering","volume":"31","author":"Gaj","year":"2013","journal-title":"Trends Biotechnol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1146\/annurev-biophys-062215-010822","article-title":"CRISPR\u2014Cas9 Structures and Mechanisms","volume":"46","author":"Jiang","year":"2017","journal-title":"Annu. Rev. Biophys."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/B978-0-12-801185-0.00008-8","article-title":"Genome editing using Cas9 nickases","volume":"546","author":"Trevino","year":"2014","journal-title":"Methods Enzymol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"foy039","DOI":"10.1093\/femsyr\/foy039","article-title":"Design principles for nuclease-deficient CRISPR-based transcriptional regulators","volume":"18","author":"Jensen","year":"2018","journal-title":"FEMS Yeast Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1016\/j.bbagrm.2012.01.002","article-title":"Chromatin dynamics in DNA double-strand break repair","volume":"1819","author":"Shi","year":"2012","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"691","DOI":"10.3389\/fgene.2018.00691","article-title":"Methodologies for Improving HDR Efficiency","volume":"9","author":"Liu","year":"2019","journal-title":"Front. Genet."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e1500454","DOI":"10.1126\/sciadv.1500454","article-title":"CRISPR-Cas9 delivery to hard-to-transfect cells via membrane deformation","volume":"1","author":"Han","year":"2015","journal-title":"Sci. Adv."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1089\/hum.2016.009","article-title":"CRISPRs for Optimal Targeting: Delivery of CRISPR Components as DNA, RNA, and Protein into Cultured Cells and Single-Cell Embryos","volume":"27","author":"Kouranova","year":"2016","journal-title":"Hum. Gene Ther."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e1602814","DOI":"10.1126\/sciadv.1602814","article-title":"CRISPR-Cpf1 correction of muscular dystrophy mutations in human cardiomyocytes and mice","volume":"3","author":"Zhang","year":"2017","journal-title":"Sci. Adv."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Sanz, D.J., Hollywood, J.A., Scallan, M.F., and Harrison, P.T. (2017). Cas9\/gRNA targeted excision of cystic fibrosis-causing deep-intronic splicing mutations restores normal splicing of CFTR mRNA. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0184009"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"26027","DOI":"10.18632\/oncotarget.15215","article-title":"The CRISPR\/Cas9 system efficiently reverts the tumorigenic ability of BCR\/ABL in vitro and in a xenograft model of chronic myeloid leukemia","volume":"8","author":"Benito","year":"2017","journal-title":"Oncotarget"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1038\/s41591-019-0500-9","article-title":"Allele-specific gene editing prevents deafness in a model of dominant progressive hearing loss","volume":"25","author":"Pan","year":"2019","journal-title":"Nat. Med."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Diakatou, M., Manes, G., Bocquet, B., Meunier, I., and Kalatzis, V. (2019). Genome Editing as a Treatment for the Most Prevalent Causative Genes of Autosomal Dominant Retinitis Pigmentosa. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20102542"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Anzalone, A.V., Randolph, P.B., Davis, J.R., Sousa, A.A., Koblan, L.W., Levy, J.M., Chen, P.J., Wilson, C., Newby, G.A., and Raguram, A. (2019). Search-and-replace genome editing without double-strand breaks or donor DNA. Nature.","DOI":"10.1038\/s41586-019-1711-4"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"e00017","DOI":"10.1128\/JVI.00017-19","article-title":"Glucosylceramidase Maintains Influenza Virus Infection by Regulating Endocytosis","volume":"93","author":"Drews","year":"2019","journal-title":"J. Virol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1851","DOI":"10.1194\/jlr.RA119000154","article-title":"Role of \u03b2-glucosidase 2 in aberrant glycosphingolipid metabolism: Model of glucocerebrosidase deficiency in zebrafish","volume":"60","author":"Lelieveld","year":"2019","journal-title":"J. Lipid Res."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Song, H.-Y., Chiang, H.-C., Tseng, W.-L., Wu, P., Chien, C.-S., Leu, H.-B., Yang, Y.-P., Wang, M.-L., Jong, Y.-J., and Chen, C.-H. (2016). Using CRISPR\/Cas9-Mediated GLA Gene Knockout as an In Vitro Drug Screening Model for Fabry Disease. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17122089"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1136\/jmedgenet-2019-106005","article-title":"Mutation-specific Fabry disease patient-derived cell model to evaluate the amenability to chaperone therapy","volume":"56","author":"Lenders","year":"2019","journal-title":"J. Med. Genet."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Song, H.Y., Chien, C.S., Yarmishyn, AA., Chou, S.J., Yang, Y.P., Wang, M.L., Wang, C.Y., Leu, H.B., Yu, W.C., and Chang, Y.L. (2019). Generation of GLA-Knockout Human Embryonic Stem Cell Lines to Model Autophagic Dysfunction and Exosome Secretion in Fabry Disease-Associated Hypertrophic Cardiomyopathy. Cells, 8.","DOI":"10.3390\/cells8040327"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"F1015","DOI":"10.1152\/ajprenal.00283.2016","article-title":"Characterization and phosphoproteomic analysis of a human immortalized podocyte model of Fabry disease generated using CRISPR\/Cas9 technology","volume":"311","author":"Pereira","year":"2016","journal-title":"Am. J. Physiol. Physiol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.kint.2018.08.033","article-title":"\u03b1-galactosidase A deficiency promotes von Willebrand factor secretion in models of Fabry disease","volume":"95","author":"Kang","year":"2019","journal-title":"Kidney Int."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"e99171","DOI":"10.1172\/jci.insight.99171","article-title":"Neuropathic pain in a Fabry disease rat model","volume":"3","author":"Miller","year":"2018","journal-title":"JCI Insight"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"AB043","DOI":"10.21037\/atm.2017.s043","article-title":"AB043. Correction of the GLA IVS4 + 919 G > A mutation with CRISPR\/Cas9 deletion strategy in fibroblasts of Fabry disease","volume":"5","author":"Chang","year":"2017","journal-title":"Ann. Transl. Med"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"15057","DOI":"10.1038\/mtm.2015.57","article-title":"Construction of a hybrid \u03b2-hexosaminidase subunit capable of forming stable homodimers that hydrolyze GM2 ganglioside in vivo","volume":"3","author":"Tropak","year":"2016","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1194\/jlr.M081323","article-title":"Cerebral organoids derived from Sandhoff disease-induced pluripotent stem cells exhibit impaired neurodifferentiation","volume":"59","author":"Allende","year":"2018","journal-title":"J. Lipid Res."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1007\/978-1-4939-6875-6_7","article-title":"CRISPR\/Cas9-Mediated Generation of Niemann\u2013Pick C1 Knockout Cell Line","volume":"1583","author":"Du","year":"2017","journal-title":"Methods Mol. Biol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"dmm034165","DOI":"10.1242\/dmm.034165","article-title":"Modeling Niemann-Pick disease type C1 in zebrafish: A robust platform for in vivo screening of candidate therapeutic compounds","volume":"11","author":"Tseng","year":"2018","journal-title":"Dis. Model. Mech."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1515\/hsz-2018-0118","article-title":"Model construction of Niemann-Pick type C disease in zebrafish","volume":"399","author":"Lin","year":"2018","journal-title":"Biol. Chem."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1016\/j.isci.2019.04.036","article-title":"CRISPR\/Cas9 Genome Engineering in Engraftable Human Brain-Derived Neural Stem Cells","volume":"15","author":"Dever","year":"2019","journal-title":"iScience"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1016\/j.bbadis.2014.01.008","article-title":"Molecular cloning and knockdown of galactocerebrosidase in zebrafish: New insights into the pathogenesis of Krabbe\u2019s disease","volume":"1842","author":"Zizioli","year":"2014","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_69","first-page":"100513","article-title":"Human GLB1 knockout cerebral organoids: A model system for testing AAV9-mediated GLB1 gene therapy for reducing GM1 ganglioside storage in GM1 gangliosidosis","volume":"21","author":"Latour","year":"2019","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.ymgme.2018.11.002","article-title":"Comprehensive behavioral and biochemical outcomes of novel murine models of GM1-gangliosidosis and Morquio syndrome type B","volume":"126","author":"Przybilla","year":"2019","journal-title":"Mol. Genet. Metab."},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Martinez-Lage, M., Puig-Serra, P., Menendez, P., Torres-Ruiz, R., and Rodriguez-Perales, S. (2018). CRISPR\/Cas9 for Cancer Therapy: Hopes and Challenges. Biomedicines, 6.","DOI":"10.3390\/biomedicines6040105"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"e349","DOI":"10.1038\/mtna.2016.58","article-title":"CRISPR-Cas9 for in vivo Gene Therapy: Promise and Hurdles","volume":"5","author":"Dai","year":"2016","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1007\/s10529-018-2555-y","article-title":"Therapeutic applications of CRISPR\/Cas9 system in gene therapy","volume":"40","author":"Mollanoori","year":"2018","journal-title":"Biotechnol. Lett."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"4482","DOI":"10.3390\/v7082830","article-title":"Special Issue: Gene Therapy with Emphasis on RNA Interference","volume":"7","author":"Lundstrom","year":"2015","journal-title":"Viruses"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1002\/path.1896","article-title":"Viral gene therapy strategies: From basic science to clinical application","volume":"208","author":"Young","year":"2006","journal-title":"J. Pathol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/23\/5897\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:37:08Z","timestamp":1760189828000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/23\/5897"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,11,24]]},"references-count":75,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["ijms20235897"],"URL":"https:\/\/doi.org\/10.3390\/ijms20235897","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,11,24]]}}}