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Engineering and characterization of a mutant ””PbKU70 for the human pathogenic fungus Paracoccidioides brasiliensis

Grant number: 15/09727-6
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): August 01, 2015
Effective date (End): February 28, 2019
Field of knowledge:Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Wagner Luiz Batista
Grantee:Alison Felipe Alencar Chaves
Host Institution: Instituto de Ciências Ambientais, Químicas e Farmacêuticas (ICAQF). Universidade Federal de São Paulo (UNIFESP). Campus Diadema. Diadema , SP, Brazil


Thermal dimorphic fungi from genera Paracoccidioides are causers of Paracoccidioidomycosis (PCM), a disease that affect mainly workers from rural area. Recent reports has shed light into the virulence factors from the fungus by using antisense RNA technology. Although important, the silencing of a gene does not ensure the causal role from that gene through the observed phenotype. To establish a causal relationship between the gene and its respective phenotype, we should avoid completely the expression of the gene product and then observe the phenotype acquired. Unfortunately, conventional gene knockout strategies are few effective when applied to fungi from Paracoccidioides genera. A strategy that has become valuable to solve similar problems in other microorganisms is the induction of homologous integration by the suppressed expression of the gene KU70 and/or Ku80 involved in the non-homologous end-joining pathway to repair the DNA. Mutants with higher rates of homologous integration has been acquired by using this procedure. At the Paracoccidioides brasiliensis genome database there is annotation to Ku70 gene, which makes this gene a potential target. Therefore, this research aims to suppress the KU70 gene from P. brasiliensis in order to establish a mutant strain whose phenotype allows anyone perform gene knockout. The possibility of perform knockouts at P. brasiliensis fungus will allow functional studies of genes responsible to code virulence factors or resistance to several types of stress inflicted by host cells. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
CHAVES, ALISON F. A.; NAVARRO, MARINA V.; CASTILHO, DANIELE G.; CALADO, JULIANA C. P.; CONCEICAO, PALLOMA M.; BATISTA, WAGNER L.. A conserved dimorphism-regulating histidine kinase controls the dimorphic switching in Paracoccidioides brasiliensis. FEMS Yeast Research, v. 16, n. 5, . (14/13961-1, 15/09727-6)
CHAVES, ALISON FELIPE ALENCAR; XANDER, PATRICIA; ROMERA, LAVINIA MARIA DAL'MAS; FONSECA, FERNANDO LUIZ AFFONSO; BATISTA, WAGNER LUIZ. What is the elephant in the room when considering new therapies for fungal diseases?. CRITICAL REVIEWS IN MICROBIOLOGY, v. 47, n. 3, . (15/09727-6, 17/04592-0)

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