Title:	Engineered fluorescent strains of cryptococcus neoformans : a versatile toolbox for studies of host-pathogen interactions and fungal biology, including the viable but nonculturable state
Authors:	Castro, Raffael Júnio Araújo de Rêgo, Marco Túlio Aidar Mariano Brandão, Fabiana S. Alfonso Pérez, Ana Laura De Marco, Janice Lisboa Fonseca, Marcio José Poças Nichols, Connie Alspaugh, James Andrew Felipe, Maria Sueli Soares Alanio, Alexandre Bocca, Anamélia Lorenzetti Fernandes, Larissa
metadata.dc.contributor.email:	mailto:larissaf@unb.br
metadata.dc.identifier.orcid:	https://orcid.org/0000-0002-5207-8892 https://orcid.org/0000-0003-3009-627X https://orcid.org/0000-0001-9726-3082 https://orcid.org/0000-0003-3323-5300 https://orcid.org/0000-0002-2814-7501
Assunto::	Cryptococcus neoformans Fungos patogênicos Cepas fluorescentes
Issue Date:	25-Aug-2022
Citation:	CASTRO, Raffael Júnio Araújo de et. al. Engineered fluorescent strains of cryptococcus neoformans: a versatile toolbox for studies of host-pathogen interactions and fungal biology, including the viable but nonculturable state. Microbiology Spectrum, v. 10, n. 5, out. 2022. DOI 10.1128/spectrum.01504-22. Disponível em: https://journals.asm.org/doi/10.1128/spectrum.01504-22. Acesso em: 28 nov. 2022.
Abstract:	Cryptococcus neoformans is an opportunistic fungal pathogen known for its remarkable ability to infect and subvert phagocytes. This ability provides survival and persistence within the host and relies on phenotypic plasticity. The viable but nonculturable (VBNC) phenotype was recently described in C. neoformans, whose study is promising in understanding the pathophysiology of cryptococcosis. The use of fluorescent strains is improving host interaction research, but it is still underexploited. Here, we fused histone H3 or the poly(A) binding protein (Pab) to enhanced green fluorescent protein (eGFP) or mCherry, obtaining a set of C. neoformans transformants with different colors, patterns of fluorescence, and selective markers (hygromycin B resistance [Hygr ] or neomycin resistance [Neor]). We validated their similarity to the parental strain in the stress response, the expression of virulence-related phenotypes, mating, virulence in Galleria mellonella, and survival within murine macrophages. PAB-GFP, the brightest transformant, was successfully applied for the analysis of phagocytosis by flow cytometry and fluorescence microscopy. Moreover, we demonstrated that an engineered fluorescent strain of C. neoformans was able to generate VBNC cells. GFP-tagged Pab1, a key regulator of the stress response, evidenced nuclear retention of Pab1 and the assembly of cytoplasmic stress granules, unveiling posttranscriptional mechanisms associated with dormant C. neoformans cells. Our results support that the PAB-GFP strain is a useful tool for research on C. neoformans.
metadata.dc.description.unidade:	Faculdade UnB Ceilândia (FCE) Colegiado de Bases Biológicas e da Saúde (FCE-BASES)
Licença::	Copyright © 2022 de Castro et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Fonte: https://journals.asm.org/doi/pdf/10.1128/spectrum.01504-22. Acesso em: 28 nov. 2022.
DOI:	https://doi.org/10.1128/spectrum.01504-22
Appears in Collections:	Artigos publicados em periódicos e afins UnB - Professores Eméritos

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