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Mechanisms involved in the generation of chromosome rearrangements and aneuploidy in Trypanosoma cruzi exposed to ionizing radiation

Grant number: 17/23312-9
Support Opportunities:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): December 01, 2017
Effective date (End): July 31, 2022
Field of knowledge:Biological Sciences - Parasitology - Protozoology of Parasites
Principal Investigator:José Franco da Silveira Filho
Grantee:Fernanda Sycko Uliana Marchiano
Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
Associated research grant:16/15000-4 - Trypanosoma cruzi: intra and interspecific genomic variability and mechanisms of cell invasion/egress, AP.TEM


The induction of DSB (Double Strand Break) on the double strand of DNA by means of ionizing radiation is a useful tool to investigate genetic recombination events in eukaryotes. Mechanisms of repair by DSB contribute to genomic plasticity and may play an important role in chromosomal polymorphism and rearrangements in fungi and protozoa. Our objective is to study the repair of DSB induced by ionizing radiation in T. cruzi and its role in generating genetic diversity observed in different strains and strains of the parasite. In addition, we will investigate the effects of ionizing radiation on T. cruzi virulence in mice. We follow the steps below for achieving these goals: 1) to analyze the effects of ionizing radiation on the molecular karyotype and gene synteny in T. cruzi by hybridization of chromosomal bands separated by Pulsed Field Gel Electrophoresis (PFGE) with chromosome-specific markers. We will select clones (CL Brener) obtained by treatment of the parasite with different doses (100, 250 and 500 Gy) of gamma ionizing radiation. Karyotype changes involving chromosomal regions e100 kb (macroalterations) will be identified by this procedure; 2) to identify chromosomal changes by comparative genomic hybridization based on DNA microarrays (aCGH). aCGH will be used for global mapping and detection of genomic changes induced by gamma radiation. Clones showing chromosomal rearrangements will be examined for chromosomal ploidy (monosomy and polysomia); 3) the virulence of T. cruzi clones with chromosomal rearrangements will be investigated by infection of mice. In this way we intend to obtain new information on the generation of the genetic diversity observed in the different strains and strains of the parasite. (AU)

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