{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T14:40:11Z","timestamp":1768315211695,"version":"3.49.0"},"reference-count":71,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T00:00:00Z","timestamp":1768262400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NORTE 2030 program in FEDER and FCT","award":["NORTE2030-FEDER-00717300"],"award-info":[{"award-number":["NORTE2030-FEDER-00717300"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Sporothrix species are thermally dimorphic fungi responsible for sporotrichosis, a globally prevalent subcutaneous mycosis and an emerging zoonotic threat, particularly in South America. The high virulence of Sporothrix brasiliensis and its efficient transmission from cats to humans have intensified recent outbreaks, underscoring the importance of understanding the pathogenic mechanisms. While several putative virulence factors have been identified, such as melanin production, cell wall remodeling, extracellular vesicles, and thermotolerance, functional studies remain hampered by limited molecular tools. Recent advances, including random mutagenesis, protoplast-mediated transformation, Agrobacterium tumefaciens-mediated transformation, RNA interference and CRISPR\/Cas9-based genome editing, are changing this landscape. These methods have enabled the functional validation of key virulence factors and the investigation of gene function in both environmental and clinical strains. In this review, we summarize the genetic toolbox available for Sporothrix, outline current challenges, and discuss how these strategies are reshaping the study of fungal virulence and host\u2013pathogen interactions.<\/jats:p>","DOI":"10.3390\/jof12010061","type":"journal-article","created":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T08:17:32Z","timestamp":1768292252000},"page":"61","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Genetic Manipulation in Sporothrix Species: Molecular Tools, Challenges, and Applications"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-7821-1586","authenticated-orcid":false,"given":"Mafalda","family":"Barros","sequence":"first","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4806-909 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4034-7362","authenticated-orcid":false,"given":"Matheus","family":"Tavares","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4806-909 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9270-2717","authenticated-orcid":false,"given":"Ricardo","family":"Silvestre","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4806-909 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9641-1792","authenticated-orcid":false,"given":"Roberta","family":"Peres da Silva","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Parasitology, Paulista School of Medicine, Federal University of S\u00e3o Paulo (UNIFESP), S\u00e3o Paulo 04023-062, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8436-9398","authenticated-orcid":false,"given":"Fernando","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4806-909 Braga, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/s12281-024-00500-9","article-title":"Insights from Cutting-Edge Diagnostics and Epidemiology of Sporotrichosis and Taxonomic Shifts in Sporothrix","volume":"19","author":"Machado","year":"2025","journal-title":"Curr. Fungal Infect. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1128\/CMR.00007-11","article-title":"Sporothrix schenckii and Sporotrichosis","volume":"24","author":"Barros","year":"2011","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e13806","DOI":"10.1111\/myc.13806","article-title":"Sporothrix and Sporotrichosis: A South African Perspective on a Growing Global Health Threat","volume":"67","author":"Fuchs","year":"2024","journal-title":"Mycoses"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Rodrigues, A.M., De Hoog, G.S., and De Camargo, Z.P. (2016). Sporothrix Species Causing Outbreaks in Animals and Humans Driven by Animal\u2013Animal Transmission. PLoS Pathog., 12.","DOI":"10.1371\/journal.ppat.1005638"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Xavier, M.O., Poester, V.R., Tr\u00e1paga, M.R., and Stevens, D.A. (2023). Sporothrix brasiliensis: Epidemiology, Therapy, and Recent Developments. J. Fungi, 9.","DOI":"10.3390\/jof9090921"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Poester, V.R., Xavier, M.O., Munhoz, L.S., Basso, R.P., Zancop\u00e9-Oliveira, R.M., Freitas, D.F.S., and Pasqualotto, A.C. (2024). Sporothrix Brasiliensis Causing Atypical Sporotrichosis in Brazil: A Systematic Review. J. Fungi, 10.","DOI":"10.3390\/jof10040287"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Do Monte Alves, M., Pipolo Milan, E., Da Silva-Rocha, W.P., Soares De Sena Da Costa, A., Ara\u00fajo Maciel, B., Cavalcante Vale, P.H., De Albuquerque, P.R., Lopes Lima, S., Salles De Azevedo Melo, A., and Messias Rodrigues, A. (2020). Fatal Pulmonary Sporotrichosis Caused by Sporothrix brasiliensis in Northeast Brazil. PLoS Negl. Trop. Dis., 14.","DOI":"10.1371\/journal.pntd.0008141"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Gremi\u00e3o, I.D.F., Miranda, L.H.M., Reis, E.G., Rodrigues, A.M., and Pereira, S.A. (2017). Zoonotic Epidemic of Sporotrichosis: Cat to Human Transmission. PLoS Pathog., 13.","DOI":"10.1371\/journal.ppat.1006077"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Garc\u00eda-Carnero, L.C., and Mart\u00ednez-\u00c1lvarez, J.A. (2022). Virulence Factors of Sporothrix schenckii. J. Fungi, 8.","DOI":"10.3390\/jof8030318"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"104555","DOI":"10.1016\/j.cellimm.2022.104555","article-title":"Differential Recognition and Cytokine Induction by the Peptidorhamnomannan from Sporothrix brasiliensis and S. schenckii","volume":"378","author":"Kischkel","year":"2022","journal-title":"Cell. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"G\u00f3mez-Gaviria, M., Mart\u00ednez-\u00c1lvarez, J.A., and Mora-Montes, H.M. (2023). Current Progress in Sporothrix brasiliensis Basic Aspects. J. Fungi, 9.","DOI":"10.3390\/jof9050533"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3730","DOI":"10.1099\/mic.0.029439-0","article-title":"Cell Surface Expression of Adhesins for Fibronectin Correlates with Virulence in Sporothrix schenckii","volume":"155","author":"Teixeira","year":"2009","journal-title":"Microbiology"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Valle-Aviles, L., Valentin-Berrios, S., Gonzalez-Mendez, R.R., and Rodriguez-del Valle, N. (2007). Functional, Genetic and Bioinformatic Characterization of a Calcium\/Calmodulin Kinase Gene in Sporothrix schenckii. BMC Microbiol., 7.","DOI":"10.1186\/1471-2180-7-107"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Rodriguez-Caban, J., Gonzalez-Velazquez, W., Perez-Sanchez, L., Gonzalez-Mendez, R., and Valle, N.R. (2011). Calcium\/Calmodulin Kinase1 and Its Relation to Thermotolerance and HSP90 in Sporothrix schenckii: An RNAi and Yeast Two-Hybrid Study. BMC Microbiol., 11.","DOI":"10.1186\/1471-2180-11-162"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1111\/j.1462-5822.2012.01813.x","article-title":"Host Carbon Sources Modulate Cell Wall Architecture, Drug Resistance and Virulence in a Fungal Pathogen","volume":"14","author":"Ene","year":"2012","journal-title":"Cell Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Lozoya-P\u00e9rez, N.E., Clavijo-Giraldo, D.M., Mart\u00ednez-Duncker, I., Garc\u00eda-Carnero, L.C., L\u00f3pez-Ram\u00edrez, L.A., Ni\u00f1o-Vega, G.A., and Mora-Montes, H.M. (2020). Influences of the Culturing Media in the Virulence and Cell Wall of Sporothrix schenckii, Sporothrix brasiliensis, and Sporothrix Globosa. J. Fungi, 6.","DOI":"10.3390\/jof6040323"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Villalobos-Duno, H.L., Barreto, L.A., Alvarez-Aular, \u00c1., Mora-Montes, H.M., Lozoya-P\u00e9rez, N.E., Franco, B., Lopes-Bezerra, L.M., and Ni\u00f1o-Vega, G.A. (2021). Comparison of Cell Wall Polysaccharide Composition and Structure between Strains of Sporothrix schenckii and Sporothrix brasiliensis. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.726958"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6874","DOI":"10.1128\/IAI.69.11.6874-6880.2001","article-title":"Involvement of Fungal Cell Wall Components in Adhesion of Sporothrix schenckii to Human Fibronectin","volume":"69","author":"Lima","year":"2001","journal-title":"Infect. Immun."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1080\/13693780802165789","article-title":"Isolation and Some Properties of a Glycoprotein of 70 kDa (Gp70) from the Cell Wall of Sporothrix schenckii Involved in Fungal Adherence to Dermal Extracellular Matrix","volume":"47","author":"Toriello","year":"2009","journal-title":"Med. Mycol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"L\u00f3pez-Ram\u00edrez, L.A., Mart\u00ednez-\u00c1lvarez, J.A., Mart\u00ednez-Duncker, I., Lozoya-P\u00e9rez, N.E., and Mora-Montes, H.M. (2024). Silencing of Sporothrix schenckii GP70 Reveals Its Contribution to Fungal Adhesion, Virulence, and the Host\u2013Fungus Interaction. J. Fungi, 10.","DOI":"10.3390\/jof10050302"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.jprot.2014.11.013","article-title":"Immunoproteomic Analysis Reveals a Convergent Humoral Response Signature in the Sporothrix schenckii Complex","volume":"115","author":"Rodrigues","year":"2015","journal-title":"J. Proteom."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Castro, R.A., Kubitschek-Barreira, P.H., Teixeira, P.A.C., Sanches, G.F., Teixeira, M.M., Quintella, L.P., Almeida, S.R., Costa, R.O., Camargo, Z.P., and Felipe, M.S.S. (2013). Differences in Cell Morphometry, Cell Wall Topography and Gp70 Expression Correlate with the Virulence of Sporothrix brasiliensis Clinical Isolates. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0075656"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Garc\u00eda-Carnero, L.C., Salinas-Mar\u00edn, R., Lozoya-P\u00e9rez, N.E., Wrobel, K., Wrobel, K., Mart\u00ednez-Duncker, I., Ni\u00f1o-Vega, G.A., and Mora-Montes, H.M. (2021). The Heat Shock Protein 60 and Pap1 Participate in the Sporothrix schenckii-Host Interaction. J. Fungi, 7.","DOI":"10.3390\/jof7110960"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"L\u00f3pez-Ram\u00edrez, L.A., Mart\u00ednez-Duncker, I., M\u00e1rquez-M\u00e1rquez, A., Vargas-Mac\u00edas, A.P., and Mora-Montes, H.M. (2022). Silencing of ROT2, the Encoding Gene of the Endoplasmic Reticulum Glucosidase II, Affects the Cell Wall and the Sporothrix schenckii\u2013Host Interaction. J. Fungi, 8.","DOI":"10.3390\/jof8111220"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.fbr.2016.09.001","article-title":"Melanin Biosynthesis in Pathogenic Species of Sporothrix","volume":"31","author":"Rozental","year":"2017","journal-title":"Fungal Biol. Rev."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Almeida-Paes, R., Figueiredo-Carvalho, M.H.G., Brito-Santos, F., Almeida-Silva, F., Oliveira, M.M.E., and Zancop\u00e9-Oliveira, R.M. (2016). Melanins Protect Sporothrix brasiliensis and Sporothrix schenckii from the Antifungal Effects of Terbinafine. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0152796"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"8623","DOI":"10.1128\/AEM.02414-12","article-title":"Biosynthesis and Functions of a Melanoid Pigment Produced by Species of the Sporothrix Complex in the Presence of l-Tyrosine","volume":"78","author":"Frases","year":"2012","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e05078-22","DOI":"10.1128\/spectrum.05078-22","article-title":"CRISPR-Based Tools for Targeted Genetic Manipulation in Pathogenic Sporothrix Species","volume":"11","author":"Hatinguais","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3696","DOI":"10.1128\/IAI.68.6.3696-3703.2000","article-title":"Biosynthesis and Functions of Melanin in Sporothrix schenckii","volume":"68","author":"Wheeler","year":"2000","journal-title":"Infect. Immun."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"105158","DOI":"10.1016\/j.micpath.2021.105158","article-title":"Melanin of Sporothrix globosa Affects the Function of THP-1 Macrophages and Modulates the Expression of TLR2 and TLR4","volume":"159","author":"Guan","year":"2021","journal-title":"Microb. Pathog."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1038\/s41564-025-01962-4","article-title":"Characterizing Extracellular Vesicles of Human Fungal Pathogens","volume":"10","author":"Rodrigues","year":"2025","journal-title":"Nat. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1128\/EC.00318-06","article-title":"Vesicular Polysaccharide Export in Cryptococcus neoformans Is a Eukaryotic Solution to the Problem of Fungal Trans-Cell Wall Transport","volume":"6","author":"Rodrigues","year":"2007","journal-title":"Eukaryot. Cell"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1007\/s11046-021-00585-7","article-title":"Extracellular Vesicles from Sporothrix brasiliensis Yeast Cells Increases Fungicidal Activity in Macrophages","volume":"186","author":"Campos","year":"2021","journal-title":"Mycopathologia"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Ikeda, M.A.K., De Almeida, J.R.F., Jannuzzi, G.P., Cronemberger-Andrade, A., Torrecilhas, A.C.T., Moretti, N.S., Da Cunha, J.P.C., De Almeida, S.R., and Ferreira, K.S. (2018). Extracellular Vesicles from Sporothrix brasiliensis Are an Important Virulence Factor That Induce an Increase in Fungal Burden in Experimental Sporotrichosis. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.02286"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3767\/003158515X687416","article-title":"Phylogeography and Evolutionary Patterns in Sporothrix Spanning More than 14 000 Human and Animal Case Reports","volume":"35","author":"Zhang","year":"2015","journal-title":"Pers\u2014Int. Mycol. J."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Huang, M., Ma, Z., and Zhou, X. (2020). Comparative Genomic Data Provide New Insight on the Evolution of Pathogenicity in Sporothrix Species. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.565439"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e00184-16","DOI":"10.1128\/genomeA.00184-16","article-title":"Draft Genome Sequence of the Dimorphic Fungus Sporothrix pallida, a Nonpathogenic Species Belonging to Sporothrix, a Genus Containing Agents of Human and Feline Sporotrichosis","volume":"4","author":"Giosa","year":"2016","journal-title":"Genome Announc."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3292","DOI":"10.1093\/gbe\/evw230","article-title":"Whole-Genome Sequencing and in Silico Analysis of Two Strains of Sporothrix globosa","volume":"8","author":"Huang","year":"2016","journal-title":"Genome Biol. Evol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e00446-14","DOI":"10.1128\/genomeA.00446-14","article-title":"Genome Sequence of the Pathogenic Fungus Sporothrix schenckii (ATCC 58251)","volume":"2","author":"Cuomo","year":"2014","journal-title":"Genome Announc."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Teixeira, M.M., De Almeida, L.G., Kubitschek-Barreira, P., Alves, F.L., Kioshima, \u00c9.S., Abadio, A.K., Fernandes, L., Derengowski, L.S., Ferreira, K.S., and Souza, R.C. (2014). Comparative Genomics of the Major Fungal Agents of Human and Animal Sporotrichosis: Sporothrix schenckii and Sporothrix brasiliensis. BMC Genom., 15.","DOI":"10.1186\/1471-2164-15-943"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1006\/fgbi.1999.1128","article-title":"Ploidy Study in Sporothrix schenkii","volume":"27","year":"1999","journal-title":"Fungal Genet. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Ferreira, B.H., Ram\u00edrez-Prado, J.H., Neves, G.W.P., Torrado, E., Sampaio, P., Felipe, M.S.S., Vasconcelos, A.T., Goldman, G.H., Carvalho, A., and Cunha, C. (2019). Ploidy Determination in the Pathogenic Fungus Sporothrix spp. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.00284"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e04564-22","DOI":"10.1128\/spectrum.04564-22","article-title":"Development of a Versatile Toolbox for Genetic Manipulation of Sporothrix brasiliensis","volume":"11","author":"Tavares","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Rodrigues, A.M., Gon\u00e7alves, S.S., De Carvalho, J.A., Borba-Santos, L.P., Rozental, S., and Camargo, Z.P.D. (2022). Current Progress on Epidemiology, Diagnosis, and Treatment of Sporotrichosis and Their Future Trends. J. Fungi, 8.","DOI":"10.3390\/jof8080776"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1007\/7651_2014_190","article-title":"Chemical and UV Mutagenesis","volume":"Volume 1373","author":"Bose","year":"2014","journal-title":"The Genetic Manipulation of Staphylococci"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1080\/02681219880000141","article-title":"UV Irradiation Induced High Frequency of Colonial Variants with Altered Morphology in Sporothrix schenckii","volume":"36","year":"1998","journal-title":"Med. Mycol."},{"key":"ref_47","first-page":"5","article-title":"Phenotypic Characterization of the Morphological Mutant UVM9 of Sporothrix schenckii","volume":"50","author":"Sabanero","year":"2008","journal-title":"Rev. Latinoam. Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1080\/mmy.39.3.295.298","article-title":"Sporothrix schenckii Thermo-Intolerant Mutants Losing Fatal Visceral Infectivity but Retaining High Cutaneous Infectivity","volume":"39","author":"Tachibana","year":"2001","journal-title":"Med. Mycol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"4026","DOI":"10.1128\/IAI.71.7.4026-4033.2003","article-title":"Synthesis of Melanin-like Pigments by Sporothrix schenckii in Vitro and during Mammalian Infection","volume":"71","author":"Youngchim","year":"2003","journal-title":"Infect. Immun."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1007\/s42770-020-00345-7","article-title":"Sporothrix globosa Melanin Inhibits Antigen presentation by Macrophages and Enhances Deep Organ Dissemination","volume":"52","author":"Song","year":"2021","journal-title":"Braz. J. Microbiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1128\/mr.53.1.148-170.1989","article-title":"Transformation in Fungi","volume":"53","author":"Fincham","year":"1989","journal-title":"Microbiol. Rev."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1016\/j.mimet.2011.01.017","article-title":"Efficient Insertional Mutagenesis System for the Dimorphic Pathogenic Fungus Sporothrix schenckii Using Agrobacterium tumefaciens","volume":"84","author":"Zhang","year":"2011","journal-title":"J. Microbiol. Methods"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/978-1-60761-611-5_1","article-title":"Protoplast Transformation of Filamentous Fungi","volume":"Volume 638","author":"Sharon","year":"2010","journal-title":"Molecular and Cell Biology Methods for Fungi"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1016\/j.funbio.2018.07.004","article-title":"Generation of Sporothrix schenckii Mutants Expressing the Green Fluorescent Protein Suitable for the Study of Host-Fungus Interactions","volume":"122","author":"Franco","year":"2018","journal-title":"Fungal Biol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"3206","DOI":"10.1002\/j.1460-2075.1995.tb07323.x","article-title":"Trans-Kingdom T-DNA Transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae","volume":"14","author":"Bundock","year":"1995","journal-title":"EMBO J."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/s40694-017-0035-0","article-title":"A Silver Bullet in a Golden Age of Functional Genomics: The Impact of Agrobacterium-Mediated Transformation of Fungi","volume":"4","author":"Idnurm","year":"2017","journal-title":"Fungal Biol. Biotechnol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.1111\/j.1365-2958.2008.06242.x","article-title":"Systematic Functional Analysis of Calcium-signalling Proteins in the Genome of the Rice-blast Fungus, Magnaporthe oryzae, Using a High-throughput RNA-silencing System","volume":"68","author":"Nguyen","year":"2008","journal-title":"Mol. Microbiol."},{"key":"ref_58","first-page":"721","article-title":"Two-component Histidine Kinase DRK1 Is Required for Pathogenesis in Sporothrix schenckii","volume":"17","author":"Zhang","year":"2018","journal-title":"Mol. Med. Report."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"99","DOI":"10.3892\/ijmm.2012.1193","article-title":"Molecular Cloning, Characterization and Differential Expression of DRK1 in Sporothrix schenckii","volume":"31","author":"Hou","year":"2013","journal-title":"Int. J. Mol. Med."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"67","DOI":"10.2147\/IDR.S185037","article-title":"Silencing of OCH1 Unveils the Role of Sporothrix schenckii N-Linked Glycans during the Host-Fungus Interaction","volume":"12","author":"Almeida","year":"2018","journal-title":"Infect. Drug Resist."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.fgb.2005.01.002","article-title":"RNA Silencing as a Tool for Exploring Gene Function in Ascomycete Fungi","volume":"42","author":"Nakayashiki","year":"2005","journal-title":"Fungal Genet. Biol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"100139","DOI":"10.1016\/j.tcsw.2024.100139","article-title":"Sporothrix brasiliensis Gp70 Is a Cell Wall Protein Required for Adhesion, Proper Interaction with Innate Immune Cells, and Virulence","volume":"13","year":"2025","journal-title":"Cell Surf."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1038\/s41587-020-0561-9","article-title":"Genome Editing with CRISPR\u2013Cas Nucleases, Base Editors, Transposases and Prime Editors","volume":"38","author":"Anzalone","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"4336","DOI":"10.1093\/nar\/gkt135","article-title":"Genome Engineering in Saccharomyces cerevisiae Using CRISPR-Cas Systems","volume":"41","author":"DiCarlo","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Morio, F., Lombardi, L., and Butler, G. (2020). The CRISPR Toolbox in Medical Mycology: State of the Art and Perspectives. PLoS Pathog., 16.","DOI":"10.1371\/journal.ppat.1008201"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1128\/EC.5.1.207-211.2006","article-title":"The akuBKU80 Mutant Deficient for Nonhomologous End Joining Is a Powerful Tool for Analyzing Pathogenicity in Aspergillus fumigatus","volume":"5","author":"Kress","year":"2006","journal-title":"Eukaryot. Cell"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1093\/mmy\/myu057","article-title":"Functional Characterization of Sporothrix schenckii Glycosidases Involved in the N-Linked Glycosylation Pathway","volume":"53","author":"Teixeira","year":"2015","journal-title":"Med. Mycol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1016\/j.resmic.2018.02.004","article-title":"Functional Characterization of the Sporothrix schenckii Ktr4 and Ktr5, Mannosyltransferases Involved in the N-Linked Glycosylation Pathway","volume":"169","year":"2018","journal-title":"Res. Microbiol."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1016\/j.funbio.2012.06.002","article-title":"Isolation and Functional Characterization of Sporothrix schenckii ROT2, the Encoding Gene for the Endoplasmic Reticulum Glucosidase II","volume":"116","author":"Lee","year":"2012","journal-title":"Fungal Biol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/j.pep.2014.12.002","article-title":"Isolation of the GFA1 Gene Encoding Glucosamine-6-Phosphate Synthase of Sporothrix schenckii and Its Expression in Saccharomyces cerevisiae","volume":"110","author":"Milewski","year":"2015","journal-title":"Protein Expr. Purif."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1007\/978-1-0716-1720-5_4","article-title":"Design and Generation of a CRISPR Interference System for Genetic Repression and Essential Gene Analysis in the Fungal Pathogen Candida albicans","volume":"Volume 2377","author":"Zhang","year":"2022","journal-title":"Essential Genes and Genomes"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/12\/1\/61\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T08:58:22Z","timestamp":1768294702000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/12\/1\/61"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,13]]},"references-count":71,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["jof12010061"],"URL":"https:\/\/doi.org\/10.3390\/jof12010061","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,13]]}}}